GSM3904695: Aif1-/- rep3; Mus musculus; RNA-Seq
Cell monolayers were harvested by trizol (Invitrogen/Life Technologies) and total RNA was extracted according to the manufacturer's protocol. Total RNA (200 ng) was used to prepare libraries using KAPA Stranded RNA-seq kit with RiboErase as described (Kapabiosystems, KR1151-v3.15). Briefly, total RNA was subjected to rRNA depletion, and double stranded cDNA (dscDNA) was synthesized followed by fragmentation. A- tailing was done at the 3'end of the dscDNA library fragments and then subjected to library amplification followed by adapter ligation for Illumina sequencing. Control RNA was spiked in (ERCC RNA Spike-In control mixes, Life Technologies) as an internal control for the library preparation and sequencing. PCR-amplified cDNA libraries were quantified on the Agilent 2100 Bioanalyzer and diluted to 10 pM for cluster generation and sequencing.