Protocol: DebM Prep 2021
DOI: 10.1000/proto.124
Protocol Source: https://protocols.io/view/library-prep-1
Pacific Biosciences SMRTbell libraries were constructed using the Revio HiFi prep kit, according to the manufacturers' instructions. DNA sequencing was performed by the Scientific Operations core at the WSI on a Pacific Biosciences Revio instrument.
For Hi-C library preparation, DNA was fragmented to a size of 400 to 600 bp using a Covaris E220 sonicator. The DNA was then enriched, barcoded, and amplified using the NEBNext Ultra II DNA Library Prep Kit following manufacturers' instructions. The Hi-C sequencing was performed using paired-end sequencing with a read length of 150 bp on an Illumina NovaSeq 6000 instrument.
Poly(A) RNA-Seq libraries were constructed using the NEB Ultra II RNA Library Prep kit, following the manufacturer's instructions. RNA sequencing was performed on the Illumina NovaSeq 6000 instrument.