Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Bisulfite-Seq
Antigen
Bisulfite-Seq

Cell type

Cell type Class
Spleen
Cell type
Spleen
MeSH Description
An encapsulated lymphatic organ through which venous blood filters.

Attributes by original data submitter

Sample

source_name
Tet2/3-DKO_24h_CMSIP
strain background
C57BL/6J
age
8-12 weeks
genotype/variation
Tet2/3-DKO (CreERT2, Tet2-flox, Tet3-flox, Rosa26-LSL-YFP)
tissue
Spleen
molecule subtype
Bisulfite-treated genomic DNA

Sequenced DNA Library

library_strategy
Bisulfite-Seq
library_source
GENOMIC
library_selection
RANDOM
library_construction_protocol
Genomic DNA isolated from naïve and activated B cells was spiked with unmethylated lambda phage cI857 Sam7 DNA (Promega, Madison, WI) and a PCR amplicon from puromycin resistant gene at a ratio of 200:1 and 100,000:1, respectively. DNA (5-10 ug in 130 uL TE buffer) was sheared with a Covaris E220 (Covaris) using microTUBE for 4 mins. DNA was cleaned-up with AmureXP beads, processed with NEBNext End Repair and A-tail Modules (NEB, Ipswich, MA), and ligated to methylated Illumina adaptors (NEB). DNA was then bisulfite-treated, denatured, and immunoprecipitated with anti-CMS serum (in house) and mixture of protein A and G dynabeads (ThermoFisher). Libraries for immunoprecipitated DNA were generated by PCR with barcoded primers (NEBNext Multiplex Oligos for Illumina, NEB) for 15 cycles using KAPA HiFi HotStart Uracil+ ReadyMix (Roche), followed by a cleanup with AmpureXP beads (Beckman Coulter). Bisulfite-seq, 50bp paired-end with HiSeq 2500

Sequencing Platform

instrument_model
Illumina HiSeq 2500

mm10

Number of total reads
22875054
Reads aligned (%)
74.8
Coverage rate (×)
0.4
Number of hyper MRs
295283 (qval < 1E-05)

mm9

Number of total reads
22875054
Reads aligned (%)
75.8
Coverage rate (×)
0.5
Number of hyper MRs
295068 (qval < 1E-05)

Base call quality data from DBCLS SRA