GSM4906366: siZNF180 transfected, rep2; Homo sapiens; RNA-Seq
Total RNA was extracted using miRNeasy QIAGEN mini kit according to the manufacturer's protocol. 500 ng of RNA were reverse transcribed using Taqman RT reagents (Applied Biosystems) with random hexamers following manufacturer's recommendations. Transcripts were quantified by real-time qPCR using Power SYBR Green PCR MasterMix (Applied Biosystems) in StepOne System (ABI). Expression levels are calculated relative to housekeeping gene, GAPDH. Total RNA was extracted 72 hr post transfection of siNTC, siZNF180, siZNF347 or siPPP1R2 in SKmel147 cell line in 3 replicates. RNA was extracted using miRNeasy QIAGEN mini kit. RNA was then processed with Ribo-Zero rRNA Removal Kit (Illumina) to remove rRNA, and further processed into sequencing libraries using Illumina TruSeq Stranded Total RNA Library Prep, following manufacturer's protocol. All libraries were sequenced on Illumina HiSeq2500 ($150M, 50 bp paired-end).