title: GSM1923426: Bulk_RNA-seq_NKT1_2; Mus musculus; RNA-Seq
library_construction_protocol: Bulk RNA-seq was performed as described in PMID:24997565 and PMID:25092889. Total RNA was purified using a miRNAeasy kit (Qiagen) and quantified as described previously (Seumois et al.Am J Clin Exp Immunol. 2012; PMID:23304658). Bulk RNA-seq was performed as described in PMID:24997565 and PMID:25092889. Briefly, 10-15 ng of purified total RNA was used for poly-A mRNA selection (Poly(A) Purist Mag kit, Life Technologies). Poly(A)-selected mRNA was amplified with a whole transcriptome amplification sequencing technology kit (SEQR, Sigma-Aldrich) as per the manufacturer’s recommendations. One microgram of this amplified cDNA was treated with the restriction enzyme (SEQR, Sigma-Aldrich) to remove the primer sequences and then purified using AMPure XP beads (Beckman Coulter). The efficiency of removal of SEQR primer sequences was assessed by quantitative PCR. 250 ng of purified amplifed cDNA was sonicated with a Covartis E220 multiplex sonicator to generate 100-250-bp DNA fragments. 250 ng DNA was used for preparing a standard SOLiD sequencing library (5500 SOLiD fragment 48 library core kit andfragment library barcode adaptors 1–96). Whole Transcriptome Amplification (MDA); cDNA amplified RNA-Seq: Following emulsion PCR (SOLiD EZ Bead E120 system kits) samples were sequenced on the 5500 SOLiD sequencer to obtain 35-bp single end reads.
attributes.tag: GEO Accession
attributes.value: GSM1923426