GSM9671114: HighFidelity_nuclear_amplicon_1; Homo sapiens; RNA-Seq
For amplicon sequencing total RNA was extracted from either whole cells or nuclear fraction For amplicon sequencing, RNA was reverse-transcribed into cDNA using the High-Capacity cDNA Reverse Transcription Kit. PCR reactions were carried out with Phire Hot Start II PCR Master Mix using 35 amplification cycles with an annealing temperature of 56 °C and a 15 s extension step. Amplified products were assessed on 2% agarose gels. Selected amplicons were submitted to Genewiz  for Amplicon-EZ next-generation sequencing and sequenced on an Illumina MiSeq platform using a 500-cycle paired-end configuration