GSM4100790: HEK293 rep-2; Homo sapiens; RNA-Seq
Cells were lysed in situ 48 hours after transfection and total RNA was isolated using miRNeasy mini kit with the optional DNase digestion step (Qiagen, Germantown MD) following the manufacturer's protocol. Using 285-400 ng RNA, cDNA was synthesized with QuantiTect Reverse Transcription kit (Qiagen, Germantown MD) following the manufacturer's protocol. cDNA was PCR amplified using barcoded primers. 2 μL cDNA were amplified in a 50 μL volume containing 2X Invitrogen Platinum SuperFi PCR Master Mix (Thermo Fisher Scientific, Waltham MA) using 1 μM (final concentration) barcoded primers. The sequencing library was created by end-polishing the barcoded PCR products, followed by adapter ligation and amplification, according to instructions of Illumina (Illumina, Inc., San Diego, CA).
GEO Accession
GSM4100790