GSM2668081: dCas9 3a3l delta_1; Homo sapiens; RNA-Seq
RNA was extracted as above, before mRNA was isolated from 1.5 µg of total RNA using mRNA DIRECT (Thermo Fisher Scientific) and fragmented with RNA fragmentation reagent (Thermo Fisher Scientific). First strand cDNA synthesis was performed with SuperScript III First-Strand Synthesis System and 3 μg/μl random hexamers (Thermo Fisher Scientific) followed by second strand synthesis with DNA polymerase I and RNase H. After purification, the double stranded cDNA was ligated to in house designed adapters (based on TruSeq Indexed adapters (Illumina)) using NEBNext Ultra II (NEB) followed by 15 cycles of amplification and library purification.
GEO Accession
GSM2668081