RNA-Seq of LCLs from the 1000 Genomes Project: NA20507
Frozen cell pellets ( 5 million cells per sample) were recovered by Coriell and cultured for 4 days (see Coriell LCL culture FAQ: https://www.coriell.org/0/sections/support/global/Lymphoblastoid.aspx). After growth, cells were transferred to a growth-limiting shipping media and were shipped directly to GENEWIZ (same-day delivery) for RNA isolation, library prep, and sequencing. Cells were spun down, then total RNA was extracted from cell pellets using Qiagen RNeasy Plus Universal mini kit following manufacturers instructions. The RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using manufacturers instructions, and the polyA enrichment workflow. Sequencing libraries were validated on the Agilent TapeStation and quantified by using Qubit 2.0 Fluorometer as well as by quantitative PCR. Sequencing libraries were multiplexed in batches of 48 samples and loaded on the flow cell on the Illumina NovaSeq 6000 instrument according to manufacturers instructions. Samples were sequenced using a 2x150 Pair-End configuration.