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16s rDNA tagging

16s rDNA tagging ?

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  • Metagenomics
  • Microorganism classification
  • Microorganism population study
  • Species identification

Metagenomic survey vs. 16s rDNA tagging

Sequencing
target
Sequencing
methods
Data
16s rRNA
tagging
16s rRNA
hypervariable
regions
Roche 454
1.Taxonomic classification
2.OTU analysis
Whole genome
metagenomic
survey
Whole
bacterial
genomes
Illumina HiSeq
1.Species identification
2.Gene prediction
3.ORF/functional annotation

16s rDNA tagging - ½ÇÇè ÁøÇà °úÁ¤

Sample requirements

concentration Concentration (ng/§¡) Quality
Genomic DNA > 5§¶ (in general) > 50 OD(260/280)>1.8 is highly recommended

Sequencing Strategy

91 ~ 101 PE sequencing

Bioinformatic analysis - contents

contents
1Â÷ ºÐ¼®
1. Illumina QC report
2. Remove the low-quality reads (poly-ATGC) and adaptors
3. Overlap the paired-end reads to form tags
4. Remove the primer
2Â÷ ºÐ¼® 1. Species classification analysis by BlastN, OTU analysis
2. Calculate alpha diversity for complexity analysis in single sample
3. Species (or OTU) ralative abundance analysis
4. Primary comparative analysis:
5. Intermediate comparative analysis :
6. Advanced comparative analysis (Sample Groups ¡Ã 2; Samples in each Group ¡Ã 10):