AMPLICON-Seq of Rattus norvegicus: adult toe
The target region was first amplified with barcoded primers (first round PCR, PCR1) using Phanta Max Super-Fidelity DNA Polymerase (P505, Vazyme). Then the PCR1 products were pooled with equal amount and purified by gel extraction or Exonuclease I digestion to remove trace of barcoded primers, followed by the second round PCR (PCR2) using index primers (N321/N322, Vazyme). Libraries were purified using DNA Clean Beads (N486, Vazyme) for sequencing using Illumina NovaSeq platform.