Amplicon Sequencing

Targeted sequencing of amplified genomic regions to identify genetic variation and support microbial profiling, mutation analysis, and biodiversity studies.

Amplicon sequencing, encompassing short-read and long-read approaches, offers a comprehensive toolkit for advanced microbial community research. Targeted Amplicon Sequencing (16S/18S/ITS) employs next-generation sequencing (NGS) of hypervariable gene regions to accurately identify microbial species within complex samples, utilizing universal primers for conserved genes like 16S rRNA, 18S rRNA, and ITS. Meanwhile, Full-Length Amplicon Sequencing maximizes taxonomic resolution by sequencing entire conserved gene regions with high-fidelity PacBio Revio technology, enabling deep and detailed profiling of complex microbial ecosystems.

Choosing the Right Amplicon Sequencing Solution:

1. Targeted Amplicon Sequencing (16S/18S/ITS) via Illumina: A high-throughput, cost-effective option ideal for taxonomic classification and comparative analysis across large sample sets.

2. Full-Length Amplicon Sequencing (16S/18S/ITS) via PacBio: Provides strain-level resolution and high precision, making it perfect for analyzing complex microbial ecosystems with greater accuracy.

Why Choose Novogene for Amplicon Sequencing?

Plentiful Experience

Annual delivery of samples over 600k amplicon metagenomic sequencing

User-Friendly Analysis

Analyze your data securely anytime with NovoMagic, our cloud platform.

Localized Support

Regional technical teams provide personalized assistance.

Customizable Solutions

Expertly designed projects for complex samples and challenging studies.

Fast Turnaround

Receive your data in as little as two weeks after sample submission.

Top-Quality Data

Industry-leading sequencing quality, validated by Zymo standards and strict contamination controls.

Applications of Amplicon Sequencing in Research and Medicine

Discover how Amplicon sequencing can enhance your research and unlock valuable insights into microbial communities.

Easy Microbial Profiling

Identify and classify microbes in complex samples by targeting 16S/18S/ITS regions, no need to culture them.

Precise Taxonomic Analysis

Achieve fine-scale microbial classification, from phylum to species, using hypervariable gene regions.

Analyze Diversity and Abundance

Measure microbial richness, abundance, and compositional differences across samples.

Detect New Lineages

pot rare or unclassified microbes with advanced database matching, identifying new candidates.

Predict Microbial Functions

Link taxonomic profiles to functional pathways such as metabolism or resistance.

Explore Ecological Links

Understand microbial shifts associated with environmental changes or host factors.

Specifications

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Novogene provides high-quality products and expert services throughout the entire project workflow. Every step is carefully designed and executed to meet rigorous scientific standards, ensuring exceptional research outcomes. To guarantee the accuracy and reliability of sequencing data, stringent quality control (QC) measures are implemented at each stage of the process. The workflow encompasses key steps such as sample preparation and quantification, fragmentation and library preparation, library quality control, sequencing, and bioinformatics analysis.

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Specifications

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Demo Results

1/1

Relative Abundance

The Y-axis shows "Relative Abundance," while the X-axis represents "Sample Names." "Others" refers to the combined relative abundance of all phyla except the top 10.

1/1

Taxonomic Abundance Cluster Heatmap

The X-axis represents sample names, and the Y-axis represents genera. The 'z' value indicates the distance from the raw score to the mean in terms of standard deviation, with negative 'z' values indicating scores below the mean.

1/1

Venn Diagram

Each circle represents a sample or group. Overlapping areas show common feature sequences, while non-overlapping areas show unique sequences specific to each sample.

1/1

Evolutionary Tree in Genus

Branches are colored by phyla, with genus-level relative abundance shown outside the circle. Different colors represent groups.

1/1

Rarefaction Abundance Curves

Each curve represents a sample, colored and shaped by sample name. The X-axis shows sequence numbers, and the Y-axis shows observed OTUs.

1/1

UPGMA Cluster Tree

The UPGMA tree on the left is paired with a relative abundance heatmap of phyla on the right, based on Weighted Unifrac Distance.

1/1

Principal Coordinates Analysis (PCoA)

Each point represents a sample, plotted by two principal components on the X and Y axes, with color coding by group. The axes percentages show the contribution to sample disparity.

1/1

Principal Component Analysis (PCA)

Each point represents a sample, with the first and second principal components on the X and Y axes respectively, colored by group.

1/1

Cladogram of LEfSe Analysis

The histogram shows LDA scores of biomarkers (species) with significant abundance differences among groups.

Demo Results

1/1

Relative Abundance

The Y-axis shows "Relative Abundance," while the X-axis represents "Sample Names." "Others" refers to the combined relative abundance of all phyla except the top 10.

1/1

Taxonomic Abundance Cluster Heatmap

1/1

Venn Diagram

Each circle represents a sample or group. Overlapping areas show common feature sequences, while non-overlapping areas show unique sequences specific to each sample.

1/1

Evolutionary Tree in Genus

Branches are colored by phyla, with genus-level relative abundance shown outside the circle. Different colors represent groups.

1/1

Rarefaction Abundance Curves

Each curve represents a sample, colored and shaped by sample name. The X-axis shows sequence numbers, and the Y-axis shows observed OTUs.

1/1

UPGMA Cluster Tree

The UPGMA tree on the left is paired with a relative abundance heatmap of phyla on the right, based on Weighted Unifrac Distance.

1/1

Principal Coordinates Analysis (PCoA)

Each point represents a sample, plotted by two principal components on the X and Y axes, with color coding by group. The axes percentages show the contribution to sample disparity.

1/1

Principal Component Analysis (PCA)

Each point represents a sample, with the first and second principal components on the X and Y axes respectively, colored by group.

1/1

Cladogram of LEfSe Analysis

The histogram shows LDA scores of biomarkers (species) with significant abundance differences among groups.

Brochure

Metagenomic Solutions Brochure

Technical Note

Technical Note to Phased Primers

More Services

Shotgun Metagenomic Sequencing

Metatranscriptome Sequencing

Microbial Whole Genome Sequencing

Microbial De novo Sequencing

Untargeted Metabolomics

Ready to Start Your Project?

Our platform offers tailored solutions for

your unique experimental needs, ensuring a seamless experience from project design to data delivery.

Amplicon Sequencing

Targeted sequencing of amplified genomic regions to identify genetic variation and support microbial profiling, mutation analysis, and biodiversity studies.

Choosing the Right Amplicon Sequencing Solution:

1. Targeted Amplicon Sequencing (16S/18S/ITS) via Illumina: A high-throughput, cost-effective option ideal for taxonomic classification and comparative analysis across large sample sets.

2. Full-Length Amplicon Sequencing (16S/18S/ITS) via PacBio: Provides strain-level resolution and high precision, making it perfect for analyzing complex microbial ecosystems with greater accuracy.

Why Choose Novogene for Amplicon Sequencing?

Plentiful Experience

Annual delivery of samples over 600k amplicon metagenomic sequencing

User-Friendly Analysis

Analyze your data securely anytime with NovoMagic, our cloud platform.

Localized Support

Regional technical teams provide personalized assistance.

Customizable Solutions

Expertly designed projects for complex samples and challenging studies.

Fast Turnaround

Receive your data in as little as two weeks after sample submission.

Top-Quality Data

Industry-leading sequencing quality, validated by Zymo standards and strict contamination controls.

Applications of Amplicon Sequencing in Research and Medicine

Discover how Amplicon sequencing can enhance your research and unlock valuable insights into microbial communities.

Easy Microbial Profiling

Identify and classify microbes in complex samples by targeting 16S/18S/ITS regions, no need to culture them.

Precise Taxonomic Analysis

Achieve fine-scale microbial classification, from phylum to species, using hypervariable gene regions.

Analyze Diversity and Abundance

Measure microbial richness, abundance, and compositional differences across samples.

Detect New Lineages

pot rare or unclassified microbes with advanced database matching, identifying new candidates.

Predict Microbial Functions

Link taxonomic profiles to functional pathways such as metabolism or resistance.

Explore Ecological Links

Understand microbial shifts associated with environmental changes or host factors.

Specifications

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Specifications

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sample Requirements

Sample amounts are listed for reference only. Download the Sample Requirements to learn more. For detailed information, please contact us with your customized requests.

Service	Sample Type	Amount	Volume	Concentration	Purity
Amplicon Metagenomic Sequencing	Genomic DNA	≥ 200 ng	≥ 20 μL	≥ 10 ng/μL	OD260/280 = 1.8-2.0, No degradation, no contamination, no color
PacBio Full -Length 16S/18S/ITS	Genomic DNA	≥ 300 ng	≥ 30 μL	≥ 10 ng/μL

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Sequencing and Analysis

Recommended data outputs and analysis contents displayed are for reference only. For detailed information, please contact us with your customized requests.

Service	Amplicon Metagenomic Sequencing		PacBio Full -Length 16S/18S/ITS
Sequencing Platform	Illumina NovaSeq System		PacBio Revio
Data Output	50K/100K/500K raw tags		10K/20K/30K/50K/100k clean reads
Qiime1 or Qiime2	Standard Analysis	Data Quality Control OTU/ASV Clustering Taxonomic Annotation Alpha diversity analysis (rarefaction curve, species boxplot) Beta Diversity Analyses (UPGMA, PCA, PCoA, NMDS) Community Differences Analyses (Anosim, MRPP, Adonis, Simper) Statistical Analyses (T-Test, MetaGenomSeq, Wilcoxon test, Kruskal-Wallis Test, LEfSe) Function Prediction (Qiime2 Only) Community Assembly Mechanisms( beta NTI, NST, iCAMP)	Qiime2	DADA2 denoise, OTU/ASV annotation and relative abundance analysis Alpha diversity analysis (rarefaction curve, species boxplot) Beta diversity analysis (heatmap, PCoA, PCA, NMDS) Comparative analysis between groups and within group (Adonis, Anosim, t-test, MetaGenomeSeq, LEfSe) Function prediction
Qiime1 or Qiime2	Advanced Analysis	Environmental association analysis (Spearman, CCA/RDA, VPA analysis) Correlation analysis (Network analysis, Network3D)	Qiime2

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Primers applied to Amplicon Metagenomic Sequencing

Types	Amplified Region	Fragment Length	Primers	Sequences (5’- 3’)
Bacterial 16S	V4	300 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4	300 bp	806R	GGACTACHVGGGTWTCTAAT
	V3-V4	470 bp	341F	CCTAYGGGRBGCASCAG
	V3-V4	470 bp	806R	GGACTACNNGGGTATCTAAT
	V4-V5	450 bp	515F	GTGCCAGCMGCCGCGGTAA
	V4-V5	450 bp	907R	CCGTCAATTCCTTTGAGTTT
	V5-V7 (for endophytic)	435 bp	799F	AACMGGATTAGATACCCKG
	V5-V7 (for endophytic)	435 bp	1193R	ACGTCATCCCCACCTTCC
	16S Full-length (V1-V9)	~1500 bp	F	AGRGTTYGATYMTGGCTCAG
	16S Full-length (V1-V9)	~1500 bp	R	RGYTACCTTGTTACGACTT
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch519F	CAGCCGCCGCGGTAA
Archaea V4 (AKA. Novel Archaea V4)	V4-V5	415 bp	Arch915R	GTGCTCCCCCGCCAATTCCT
Fungal 18S	V4	350 bp	528F	GCGGTAATTCCAGCTCCAA
	V4	350 bp	706R	AATCCRAGAATTTCACCTCT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-A	AACCTGGTTGATCCTGCCAGT
	18S Full-length (V1-V9)	~1800 - 2000 bp	Euk-B	GATCCTTCTGCAGGTTCACCTAC
Fungal ITS	ITS1	200-400 bp	ITS5-1737F	GGAAGTAAAAGTCGTAACAAGG
	ITS1	200-400 bp	ITS2-2043R	GCTGCGTTCTTCATCGATGC
	ITS2	380 bp	ITS3-2024F	GCATCGATGAAGAACGCAGC
	ITS2	380 bp	ITS4-2409R	TCCTCCGCTTATTGATATGC
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-F	CTTGGTCATTTAGAGGAAGTAA
	ITS1-1F (for endophytic)	200-400 bp	ITS1-1F-R	GCTGCGTTCTTCATCGATGC
	ITS Full-length (1-2)	~500 - 800 bp	ITS9munngs	TACACACCGCCCGTCG
	ITS Full-length (1-2)	~500 - 800 bp	ITS4ngsUni	CCTSCSCTTANTDATATGC

Project Workflow

Demo Results

1/1

Relative Abundance

The Y-axis shows "Relative Abundance," while the X-axis represents "Sample Names." "Others" refers to the combined relative abundance of all phyla except the top 10.

1/1

Taxonomic Abundance Cluster Heatmap

1/1

Venn Diagram

Each circle represents a sample or group. Overlapping areas show common feature sequences, while non-overlapping areas show unique sequences specific to each sample.

1/1

Evolutionary Tree in Genus

Branches are colored by phyla, with genus-level relative abundance shown outside the circle. Different colors represent groups.

1/1

Rarefaction Abundance Curves

Each curve represents a sample, colored and shaped by sample name. The X-axis shows sequence numbers, and the Y-axis shows observed OTUs.

1/1

UPGMA Cluster Tree

The UPGMA tree on the left is paired with a relative abundance heatmap of phyla on the right, based on Weighted Unifrac Distance.

1/1

Principal Coordinates Analysis (PCoA)

Each point represents a sample, plotted by two principal components on the X and Y axes, with color coding by group. The axes percentages show the contribution to sample disparity.

1/1

Principal Component Analysis (PCA)

Each point represents a sample, with the first and second principal components on the X and Y axes respectively, colored by group.

1/1

Cladogram of LEfSe Analysis

The histogram shows LDA scores of biomarkers (species) with significant abundance differences among groups.

Demo Results

1/1

Relative Abundance

The Y-axis shows "Relative Abundance," while the X-axis represents "Sample Names." "Others" refers to the combined relative abundance of all phyla except the top 10.

1/1

Taxonomic Abundance Cluster Heatmap

1/1

Venn Diagram

Each circle represents a sample or group. Overlapping areas show common feature sequences, while non-overlapping areas show unique sequences specific to each sample.

1/1

Evolutionary Tree in Genus

Branches are colored by phyla, with genus-level relative abundance shown outside the circle. Different colors represent groups.

1/1

Rarefaction Abundance Curves

Each curve represents a sample, colored and shaped by sample name. The X-axis shows sequence numbers, and the Y-axis shows observed OTUs.

1/1

UPGMA Cluster Tree

The UPGMA tree on the left is paired with a relative abundance heatmap of phyla on the right, based on Weighted Unifrac Distance.

1/1

Principal Coordinates Analysis (PCoA)

Each point represents a sample, plotted by two principal components on the X and Y axes, with color coding by group. The axes percentages show the contribution to sample disparity.

1/1

Principal Component Analysis (PCA)

Each point represents a sample, with the first and second principal components on the X and Y axes respectively, colored by group.

1/1

Cladogram of LEfSe Analysis

The histogram shows LDA scores of biomarkers (species) with significant abundance differences among groups.

Brochure

Metagenomic Solutions Brochure

Technical Note

Technical Note to Phased Primers

More Services

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Metatranscriptome Sequencing

Microbial Whole Genome Sequencing

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