GSM2335533: ASCL1 ChIPseq doxcycline rep3; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
TFs and others
Cell type Class
Attributes by original data submitter
derived cell line
Sequenced DNA Library
Lysates were clarified from sonicated nuclei and ASCL1-DNA complexes were isolated with antibody. Libraries were prepared according to NEBNext ChIP-Seq library preparation protocol. Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were selected with Ampure beads. Library was validated by Bioanalzyer and Kapa qPCR assay and sequenced on an Illumina HiSeq 2500 platform, using a Rapid Run mode flowcell to generate paired end reads of 100 bases.