Rabbit polyclonal serum against SA1 was obtained by using as immunogen a synthetic peptide (CEDDSGFGMPMF)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were cross-linked with 1% formaldehyde added to the media for 15 minutes at RT. After quenching with 0.125M Glycine, fixed cells were washed twice with PBS, pelleted and lysed in lysis buffer (1%SDS, 10mM EDTA, 50mM Tris-HCl pH8.1) at 2x107 cells/ml. Sonication was performed with a Covaris system (shearing time 20 min, 20% duty cycle, intensity 6, 200 cycles per burst and 30s per cycle) in a minimum volume of 2ml. 1 to 5ng of DNA per sample were used as provided by the user. Samples were processed through subsequent enzymatic treatments of end-repair, dA-tailing, and ligation to adapters with "NEBNext Ultra II DNA Library Prep Kit for Illumina" from New England BioLabs (catalog # E7645). Adapter-ligated libraries were completed by limited-cycle PCR and extracted with a [single] double-sided SPRI size selection. Resulting average fragment size is 370 bp from which 120 bp correspond to adaptor sequences. Libraries were applied to an Illumina flow cell for cluster generation and sequenced on an Illumina instrument (see below) by following manufacturer's protocols.