Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K4me3

Cell type

Cell type Class
Lung
Cell type
A549
Primary Tissue
Lung
Tissue Diagnosis
Carcinoma

Attributes by original data submitter

Sample

source_name
cell line not treated with flavopiridol
cell line
A549
antibody
H3K4me3

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cell or Chopped tissue pellets were homogenized in PBS containing proteinase inhibitor. The nuclei were checked by phase-contrast microscopy to maximize the yield of nuclei (~5x107). With sufficient lysis in 25% sucrose, 0.2% NP-40/TBS, the pellets were layered on 50% [w/v] sucrose/TBS, and then centrifuged at 21,000 g, 30 min, 4°C. The nuclei pellets were resuspended in digestion buffer (0.32 M sucrose, 50 mM Tris-HCl (pH 7.5), 4 mM MgCl2, 1 mM CaCl2), and digested with MNase for ~10 min to generate mainly mononucleosomes that were immunoprecipitated with anti-H3K4me3 antibody (ab8580, Abcam). After incubation over night, Protein A Dynal magnetic beads were washed 6 times using modified RIPA buffer (50 mM Tris-HCL pH 7.8, 1 mM EDTA, 0.25% Na Deoxycholate, 1% NP-40, 0.5 M LiCl). The DNA was eluted in 1% SDS and 0.1M NaHCO3 and reversed cross-linking at 65°C, over night. The library was generated using NEBNext ChIP-Seq Library Prep Master Mix Set, and amplified with 10 PCR cycles. The library was prepared by size selection for mononucleosome fragments.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

hg38

Number of total reads
58991248
Reads aligned (%)
99.4
Duplicates removed (%)
46.7
Number of peaks
64775 (qval < 1E-05)

hg19

Number of total reads
58991248
Reads aligned (%)
99.1
Duplicates removed (%)
47.1
Number of peaks
64584 (qval < 1E-05)

Base call quality data from DBCLS SRA