GSM3938205: Control_2; Mus musculus; RNA-Seq
Total RNA from sorted cells was extracted using the QIAGEN RNeasy Micro kit according to the manufacturer's protocol. cDNA synthesis was performed with SmartScribe reverse transcriptase (Clontech), universal poly-dT primers and template switching oligos. Purified cDNA was amplified with Advantage 2 DNA Polymerase in 12 cycles, and subjected to ultrasonic shearing with Covaris S2. Standard Illumina fragment libraries were prepared using NEBnext chemistries (New England Biolabs). Library preparation consisted of fragment end-repair, A-tailing and ligation to indexed Illumina TruSeq adapters, followed by a universally primed PCR amplification of 15 cycles. Then, libraries were purified using XP beads (Beckman Coulter) and quantified by qPCR.