GSM4816679: nascONT-seq K562 DMSO 2h R1(OJ48); Homo sapiens; OTHER
Cells were collected and lysed by incubating on ice in 0.15% NP-40, 25 mM α-amanitin in PBS for 2 min. The following steps of cell fractionation were performed as described by Mayer and Churchman (2016), with only minor modifications. After purification of the chromatin-associated RNA by isopropanol precipitation, and DNase treatment, the RNA way biotinylated using MTSEA biotin-XX linker (Biotinum) and purified using μMACS streptavidin MicroBeads. E. coli poly-A polymerase (Takara) was used to polyadenylate the enriched RNA. The libraries was prepared using the Direct cDNA Sequencing kit (Oxford Nanopore Technologies), with modifications to improve capture of longer fragments.
GEO Accession
GSM4816679