title: GSM8637625: mES, dTAG_SPT6, dTAG3h, rep2, PRO-seq; Mus musculus; OTHER
library_name: GSM8637625
library_construction_protocol: Nascent RNA was extracted using a Total RNA Puriﬁcation Kit following the manufacturer's instructions (ZOME). The total RNA is subjected to three biotin-Streptavidin enrichment to select nascent RNA away from other RNA. We isolated nascent RNAs with magnetic beads coated with streptavidin, followed by 3' adapter ligation. After another round of biotin-streptavidin affinity purification, we removed the mRNA cap and ligated the 5' adapter. Following the third biotin-streptavidin affinity purification, we generated cDNA by reverse transcription and prepared PRO-seq libraries for sequencing using Illumina TruSeq small-RNA adaptors with twelve cycles of PCR. The precipitated DNA samples were prepared for DNA deep sequencing according to manufacturer's guidelines (SWIFT, #21096).
identifiers.namespace: GEO
identifiers.id: GSM8637625_r1