GSM5277210: ORAI1KO HEK293-ACE2 cells - mock rep2; Homo sapiens; RNA-Seq
Samples were harvested in TriZol and RNA was extracted using the Direct-zol RNA Mini prep kit Libraries for RNA-Seq were prepared with TruSeq Stranded mRNA Library Prep Kit (Illumina). The workflow consisted of mRNA enrichment and fragmentation. Cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. Strand specificity is achieved by replacing dTTP with dUTP, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. cDNA generation is followed by A-tailing, adaptor ligation and PCR amplification. Different adaptors were used for multiplexing samples in one lane.