title: GSM4763743: CIGAR-seq of HAP1 cells; Homo sapiens; RNA-Seq
library_construction_protocol: 2x108 HAP1 cells were infected with CIGAR-seq viral particles (MOI = 0.3) and treated with 1 µg/ml of Puromycin for 24 h post infection. Puromycin resistant cells were cultured for additional seven days, and then 2x108 HAP1 cells were collected for RNA extraction. 100 ng of bisulfite-treated mRNA (see above) was quantified using Agilent RNA 6000 Pico Kit (Agilent, #NC1711873) and reverse transcribed into cDNA using HiScript II Q Select RT SuperMix (Vazyme, #R233-01). Finally, using the cDNA, CIGAR-seq NGS library was amplified with forward primer 5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-GGGTTGGTTTAGGAGATATTTGAGGG-3' and reverse primer 5'- CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-AACAATCCTAATACTCAAAAAAAAAAACACCA-3' Finally, using the cDNA, CIGAR-seq NGS library was amplified with forward primer 5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-GGGTTGGTTTAGGAGATATTTGAGGG-3' and reverse primer 5'- CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-AACAATCCTAATACTCAAAAAAAAAAACACCA-3' targeted RNA-seq
attributes.tag: GEO Accession
attributes.value: GSM4763743