GSM2443417: cx_exo_5ppp_2; Homo sapiens; RNA-Seq
Exosomes were isolated from conditioned media collected at 48-72 hours by serial high speed ultracentrifugation, or using 10% final concentration of polyethylene-glycol and low speed centrifugation. Exosome RNA was extracted with TRIzol. 0.1-2 µg of exosomal RNA was prepared by first degrading 5’monophosphate RNA with Terminator 5´-Phosphate-Dependent Exonuclease (Epicentre), then converting 5’triphosphate to 5’p with RNA 5’ Polyphosphatase (Epicentre), to allow for specific ligation of RNA adaptor P5_RNA to RNAs that originally have 5’ triphosphate. Then, cDNAs were synthesized by using a primer with 5’ random 9mer (P7_N9), and amplified with NEBNext PCR reagents (NEB).
GEO Accession
GSM2443417