title: GSM4678221: HCT116_KO_Pae1_rep2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated with the use of QIAzol Lysis (Qiagen) followed by isopropanol precipitation. Then, concentration and integrity RNAestimated with the use of Fragment Analyzer (Agilent) and Qubit 4 Fluorometer (Thermo Fisher). Poly-A tail mRNA was enriched using magnetic beads. First stand of cDNA was synthesized using SuperScript III Reverse Transcriptase (Thermo Fisher), and second strand was synthesized using the second strand master mix (TruSeq RNA Library Prep Kit V2 (Illumina)). The synthesized DNA was cleaned up with the use of AgencourtAMPure XP Beads (Beckman Coulter). After, end repair/dA-tailing, adaptors were ligated, and the library was further enriched with the use of TruSeq RNA Library Prep Kit V2 (Illumina). After determining quality and concentration, the cDNA libraries were pooled and loaded (20 pM) into the NextSeq 500 (Illumina) Sequencer with the use of NextSeq 500/550 High Output Kit v2.5(75 cycles).
attributes.tag: GEO Accession
attributes.value: GSM4678221