GSM4730318: ucMSC-TL-Seq-NegativeControl; Homo sapiens; OTHER
DECAP-seq was performed as described in ref. 18 with some modifications. The starting material was poly-A+ RNA was isolated from 100mg of total RNA using the Dynabeads mRNA Purification Kit (Life Technologies, 61006). 5' phosphate groups were removed using CIP (NEB, M0290) rather than AP, and Cap-Clip acid pyrophosphatase (CellScript, C-CC15011H) was used in place of RppH to de-cap the mRNAs. Size selection of products was performed after 5' adapter ligation, rather than after 1st strand synthesis to select against cDNAs generated solely from binding of the RT primer to the 5' adapter. A negative control library was generated by omitting the de-capping step TL-seq signal tracks were generated by bamCompare