Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Unclassified
Antigen
Unclassified

Cell type

Cell type Class
Embryonic fibroblast
Cell type
C3H/10T1/2
NA
NA

Attributes by original data submitter

Sample

source_name
C3H10T1/2 cells
cell type
C3H embryo-derived mesenchymal multipotent cells
genotype
Stably expressing FLAG-HA-tagged K36M mutant H3.3
passages
Passage 10-15

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
~2x10^7 cells were lysed using digesting buffer (50 mM Tris-HCl pH 7.6, 1 mM CaCl2 and 0.2% Triton X-100) and digested with micrococcal nuclease to obtain mono-nucleosomes. The chromatin were then dialyzed into RIPA buffer (10 mM Tris pH 7.6, 1 mM EDTA, 0.1% SDS, 0.1% Na-Deoxycholate, 1% Triton X-100) for 2 h at 4°C. After centrifugation, soluble chromatin was incubated with 3-5 μg of antibody bound to 75 μl protein A or protein G Dynal magnetic beads (Invitrogen) and incubated overnight at 4°C, with 5% kept as input DNA. Magnetic beads were washed with RIPA buffer and LiCl buffer (0.25 M LiCl, 0.5% NP40, 0.5% Na-Deoxycholate) and chromatin was eluted. ChIP DNA was treated with Proteinase K (Roche) and recovered using Qiagen PCR purification kit. libraries were prepared according to the Illumina TruSeq protocol

Sequencing Platform

instrument_model
Illumina HiSeq 2500

mm10

Number of total reads
41526926
Reads aligned (%)
98.7
Duplicates removed (%)
6.6
Number of peaks
154 (qval < 1E-05)

mm9

Number of total reads
41526926
Reads aligned (%)
98.6
Duplicates removed (%)
6.6
Number of peaks
150 (qval < 1E-05)

Base call quality data from DBCLS SRA