GSM8698870: DO mNPC Sample PB360.45; Mus musculus; RNA-Seq
Total RNA was extracted using the Quick-RNA 96 well format kit (Zymo Research) with in-column DNase treatment. Sequencing libraries were prepared by Akesogen using the TruSeq Stranded mRNA HT kit (Illumina, Cat no. 20020595) and included ribosomal RNA reduction and poly-A selection, enzymatic fragmentation, cDNA synthesis from random hexamer priming, adaptor ligation and PCR amplification steps to generate indexed, stranded mRNA-seq libraries. Libraries were checked for quality and quantitated with the Agilent Bioanalyzer, and samples that failed QC were repeated starting from the cryovial stage. Finally, pooled libraries were sequenced on the NextSeq platform (Illumina) using the NextSeq 500/550 High Output v2 150-cycle kits (Illumina, Cat no. FC-404-2002). To minimize technical variation, samples were randomly assigned to lanes prior to sample processing steps, barcoded, and multiplexed at 16 samples per flow cell, yielding 6M-55M 2x75bp paired-end (PE) reads per sample