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RNA was reverse transcribed into cDNA using the TaKaRa PrimeScript RT Master Mix kit (TaKaRa, Japan). Next, we performed amplicon sequencing spanning the whole genome of SARS-CoV-2. The primers were divided into two pools: A and B, and each included 49 pairs of primers with approximately 400bp. Singlex PCR amplification was performed using NEB Q5 Hot Start High-Fidelity 2X Master Mix (New England Biolabs, UK). Then, PCR products were purified with AMPure XP magnetic beads (Beckman, USA). The qualified libraries were sequenced on an Illumina Nova-seq Platform (Illumina, USA) using a pair-end 150-base pair (bp) strategy. |
Targeted-Capture |
VIRAL RNA |
PCR |
PAIRED
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