GSM1529691: 1.0_5azaCdR_repB; Homo sapiens; RNA-Seq
HEK 293T cells were collected by washing and trypsinizing for three minutes. The same procedure was performed with cells allowed to remain in culture for 30 days without the drug. Total RNA was extracted with Trizol (Invitrogen) and rRNA depleted with Ribozero (Epicentre). cDNA synthesis was performed by using oligo-dT as well as random hexamers.  Actinomycin D was added to the reaction to prevent any possible amplification from contaminating genomic DNA.  During second strand synthesis, a dU/VTP mix was used to create directional libraries.   Before library preparation, cDNA samples were Covaris-fragmented to 300 bp fragments.  The samples were then end-filled, 3’ terminal A-extended and ligated to pre-annealed TruSeq indexed Illumina adapters.  Uracil-DNA-glycosylase (UDG) treatment preceded the PCR reaction to cleave the uridine-containing strand and therefore identify the orientation of each transcript
GEO Accession
GSM1529691