title: GSM1563056: PC3_shCIC2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using Trizol reagent (Invitrogen, USA). For isolation of mRNA from total RNA, we used Oligotex mRNA mini kit (QIAGEN, USA). mRNA were fragmented by keeping it with 1x Superscript III reverse transcriptase buffer (Invitrogen, USA) for 4 minutes  at 95°C. The cDNA was synthesized by random hexamer and Superscript III reverse transcriptase.  Both ends of cDNA strands were filled using the Epicentre DNA END-Repair kit (Epicentre Biotechnologies, Madison, WI). Adenine was added at 3’ ends by  Taq DNA polymerase (New England Biolabs, Beverly, MA) under dATP conditions. Illumina’s adaptor oligomers was ligated at both ends using Quick Ligation kit (Qiagen, Germantown, MD). After purified twice, cDNA libraries are amplified using Solexa primers (Illumina, San Diego, CA) and Phusion PCR master mix (Thermo Scientific, Hudson, NH).
attributes.tag: GEO Accession
attributes.value: GSM1563056