anti-HA followed by anti-BrdU (B44, BD Biosciences)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Preparation of BrdU-IP, ChIP and eSPAN DNA according to a published eSPAN protocol (Yu et al., 2018). Briefly, cells were grown under the specified conditions and then fixed with 1% formaldehyde. Cells were lysed and nuclear extracts harvested, followed by chromatin immunoprecipitation. Purified ChIP DNA was then denatured and subject to another round of BrdU-IP. In parallel, BrdU-IP was also performed with Input DNA. Libraries were then amplified from purified eSPAN DNA, alongside ChIP, BrdU-IP and Input DNA, and were subject to high throughput sequencing by the Ilumina HiSeq 2500 and 4000 platforms. Sequencing libraries were prepared using the Accel-NGS® 1S Plus DNA Library Kit as per manufacturer's protocol.