Amplicon sequencing

Amplicon sequencing is frequently used to identify and differentiate microbial species. 16S rRNA gene is the most common target used for bacteria and archaea. 18S rRNA gene, and two internal transcribed spacers (ITS) located between rRNA genes are generally used for fungi.

Sequencing Strategy
1.HiSeq platform, paired-end 250 bp
2.Recommended Sequencing Data: 3W raw tags

Sample Requirements
1.DNA amount: ≥ 2 μg, DNA concentration: ≥ 50 ng/μl
2.PCR product amount: ≥ 400 ng, PCR product concentration: ≥ 20 ng/μl

Turnaround Time
40 working days

List of Analyses
1.Data quality control
2.Overlap the paired-end reads to form tags
3.OTUs clustering and filtering
4.Alpha-diversity analysis
5.OTUs analysis and species annotation( Krona results, phylogenetic composition analysis, phylogenetic tree, heatmap, taxonomic tree)
6.Beta-diversity analysis (unweighted UniFrac distance heatmap, PCA, PCoA, UPGMA, NMDS analysis)
7.Multi-variate statistical analysis( LEfSe analysis, metastats analysis, ANOSIM, and MRPP analysis)