GSM5182843: P_J1: Plasmid_replicate_Jurk_amp_1; synthetic construct; OTHER
Twenty-four hours after transfection, cells were collected and snap frozen in liquid N2. Total RNA was extracted and GFP mRNA was isolatedand converted to cDNA Tewhey and colleagues 2016. Following Tewhey et al. 2016, cDNA libraries were quanitified using qPCR including the plasmid pool used to transfect the cells. They were then diluted to be of equal concentration with the lowest library per cell type (15 cycles for K562 cells and 21.5 for Jurkat cells) and amplified using one fewer cycle than the lowest cycle threshold. An addition PCR was performed to add P7 indexing primers. Libraries were than molar pooled and sequenced  with 1x50bp chemistry on a Illumina HiSeq 2500
GEO
GSM5182843