GSM4310149: LSK cells, 14359X5 shCxxc5-2; Mus musculus; RNA-Seq
RNA was isolated from 50,000 to 20,000 sorted LSK cells using DNA/RNA isolation kit (Zymo Research- Quick-DNA/RNA Microprep Plus Kit) RNA samples were quantified using NanoDrop 8000 and Qubit RNA HS Assay. RNA integrity numbers were calculated on Agilent HS RNA ScreenTape. RNA was subjected to library construction with the Illumina TruSeq Stranded mRNA Sample Preparation Kit (Cat#RS-122-2101, RS-122-2102) according to manufacturer's protocol. Chemically denatured sequencing libraries (25 pM) are applied to an Illumina HiSeq v4 single read flow cell using an Illumina cBot. Hybridized molecules were clonally amplified and annealed to sequencing primers with reagents from an Illumina HiSeq SR Cluster Kit v4-cBot (Cat#GD-401-4001). Following the transfer of the flowcell to an Illumina HiSeq 2500 instrument (HCSv2.2.38 and RTA v1.18.61), a 50-cycle single-read or 125-cycle paired-end sequence run was performed using HiSeq SBS Kit v4 sequencing reagents (Cat#FC-401-4002).