GSM2420212: Sample_54972; Mus musculus; RNA-Seq
RNA was harvested using the RNEasy-Plus Kit (Qiagen;74136) and assessed for quality using the TapeStation (Agilent). Samples with RNA Integrity Number of 8 or greater were rRNA depleted using Ribo Minus (Invitrogen;K1550-04). rRNA-depleted samples were prepared using the TruSeq mRNA SamplePrep v2 kit (Illumina;RS-122-2001, RS-122-2002). The entire fraction of 0.1-3 ug of rRNA depleted total RNA was fragmented and copied into first-strand cDNA using reverse transcriptase and random primers. cDNA 3'-ends were adenylated and adapters were ligated. One of the adapters had a 6-nucleotide barcode that was unique for each sample, allowing us to multiplex in a HiSeq flow cell (Illumina). Products were purified and enriched by PCR to create the final cDNA library. Libraries were checked for quality and quantity by TapeStation (Agilent) and qPCR using a library quantification kit for Illumina platforms (Kapa Biosystems;KK4835). They were clustered on the cBot (illumina) and sequenced 24 samples per lane on 6 lanes of a 50-cycle single end HiSeq 4000. HiSeq Control Software version 3.3.52 was used according to manufacturer?s protocols. Demultiplexing and Fastq file generation was done using bcl2fastq version 2.17.1.14.
GEO Accession
GSM2420212