Targeted 16S rRNA Primer Design
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Corresponding Organization : Harvard University
Protocol cited in 1 other protocol
Variable analysis
- Previously obtained 16S rRNA reference sequences for lesser-known taxa in human and canine oral microbiomes
- Identification of similar sequences in Greengenes and NCBI databases
- Obtaining primer pairs that would specifically amplify 16S rDNA for lineages including known oral taxa
- Designing primer pairs with broad coverage to amplify the entire phylum or candidate division
- Manually aligning the sequences in a curated secondary structure-based rRNA database
- Selecting target regions by inspection of the alignments
- Analyzing candidate primers for melting temperature, hairpin formation, and primer dimer formation using OligoAnalyzer (Integrated DNA Technologies)
- Not explicitly mentioned
- Not explicitly mentioned
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