H > ¼ºñ½º > Next-Generation Sequencing (NGS) > 16s rDNA tagging
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): |