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The time-temperature profile for metagenomic analysis of the 16S rRNA V3-V4 hypervariable region
| NGS PCR profile | |||
|---|---|---|---|
| Step | Temperature | Time | No. of cycles |
| initial denaturation | 98 °C | 30 s | 1 |
| denaturation | 98 °C | 10 s | 3–15 |
| annealing | 65 °C | 75 s | |
| final extension | 65 °C | 5 min | 1 |
| final hold | 4–10 °C | ∞ | |
Abbreviations: 16S rRNA 16S ribosomal RNA, NGS Next generation sequencing, 16S V3-V4 Regions of interest-specific primer
