GSM2044669: RNA-seq Experimental 2; Mus musculus; RNA-Seq
Approximately 100 ng of total RNA was used for sequence library construction following instructions of NuGen mRNA sample prep kit (cat# 0348). In brief, total RNA was copied into first strand cDNA using reverse transcriptase and random primers. This was followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH. These cDNA fragments went through an end repair process, the addition of a single ‘A’ base, and then ligation of the adapters. These products were gel purified and enriched with PCR to create the final cDNA libraries. The library constructs were run on the bioanalyzer to verify the size and concentration before sequencing on the Illumina HiSeq2500 machine where 100-cycle paired-end sequencing was performed by the University of Wisconsin Biotechnology Center. In total, three libraries (N = 2 genomes/library, except N = 1 for one library; the same mice and combinations that were used to generate the 5hmC data) were sequenced for each experimental condition.
GEO Accession
GSM2044669