GSM6288608: Control4; Mus musculus; RNA-Seq
GSM6288608
Poly(A) RNA was extracted from 15,000 non-epithelial cells of an individual mouse #4 mammary glands using the Dynabeads mRNA DIRECT purification kit (Thermo Fisher, Waltham, MA) according to the manufacturer's protocol. A bulk adaptation of the MARS-Seq protocol was used to generate RNA-Seq libraries. Briefly, 30 ng of input RNA from each sample was barcoded during reverse transcription and pooled. Following Agencourct Ampure XP beads cleanup (Beckman Coulter, Indianapolis, IN), the pooled samples underwent second strand synthesis and were linearly amplified by T7 in vitro transcription. The resulting RNA was fragmented and converted into a sequencing-ready library by tagging the samples with Illumina sequences during ligation, RT, and PCR. Libraries were quantified by Qubit and TapeStation as well as by qPCR for ActB housekeeping gene.