GSM2883451: E14.5 GF Replicate 1; Mus musculus; RNA-Seq
Total RNA was extracted using the Ambion mirVana™ miRNA Isolation Kit (Ambion® Thermo Fisher Scientific, Waltham, MA, USA) according to manufacturer's protocol. cDNA libraries were prepared from 2 ng total RNA starting material and 1 ul of a 1:50,000 dilution of ERCC RNA Spike in Controls (Ambion® Thermo Fisher Scientific) using the SMARTSeq v2 protocol (Picelli et al., 2014) with the following modifications: 1. Addition of 20 µM TSO; 2. Use of 250 pg cDNA with 1/5 reaction of Illumina Nextera XT kit (Illumina, San Diego, CA, USA). OR cDNA libraries were prepared using 50 ng of total RNA and 2ul of a 1:2000 dilution of ERCC RNA Spike in Controls (Ambion). The fragmented mRNA samples were subjected to cDNA synthesis using Illumina TruSeq® RNA sample preparation kit version 2 (Low-Throughput protocol) (Illumina, San Diego, CA, USA) according to manufacturer's protocol, except for the following modifications: 1. use of 13 PCR cycles, 2. two additional round of Agencourt Ampure XP SPRI beads (Beckman Courter) to remove >600bp double stranded cDNA.
GEO Accession
GSM2883451