Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K27ac

Cell type

Cell type Class
Gonad
Cell type
Round spermatids
NA
NA

Attributes by original data submitter

Sample

source_name
Round spermatides
strain
C57BL/6
tissue
testis
age
adult
genotype
wild type
antibody
K27ac
biowardrobe ides
4318

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP-seq libraries were prepared through the ChIPmentation method (Schmidl et al., 2015). Briefly, beads were resuspended in 30 μl of the tagmentation reaction buffer (10 mM Tris-HCl pH 8.0 and 5 mM MgCl2) containing 1 μl Tagment DNA Enzyme from the Nextera DNA Sample Prep Kit (Illumina) and incubated at 37°C for 10 min in a thermocycler. The beads were washed twice with 150 μl cold wash buffer 1, incubated with elution buffer (as described above) at 42°C for 30 min, and then incubated at 65°C for another 5 h to reverse cross-linking. DNA was purified with the MinElute Reaction Cleanup Kit (Qiagen) and amplified with NEBNext High-Fidelity 2x PCR Master Mix (NEB). Amplified DNA was purified by Agencourt AMPure XP (Beckman Coulter). Afterward, DNA fragments in the 250- to 500-bp size range were prepared by agarose gel size selection. DNA libraries were adjusted to 5 nM in 10 mM Tris-HCl pH8.0 and sequenced with an Illumina HiSeq 2500. For RNA-seq libraries, total RNA was purified using an RNeasy micro kit (Qiagen) according to the manual provided. RNA quality and quantity were checked using, respectively, Bioanalyzer (Agilent) and Qubit (Life technologies). The initial amplification step was performed with the NuGEN Ovation RNA-Seq System v2. The assay was used to amplify RNA samples and create double stranded cDNA. Libraries were then created with the Nextera XT DNA Sample Preparation Kit (Illumina) and sequenced with an Illumina HiSeq 2500.

Sequencing Platform

instrument_model
Illumina HiSeq 2500

mm10

Number of total reads
17275380
Reads aligned (%)
85.5
Duplicates removed (%)
18.5
Number of peaks
11141 (qval < 1E-05)

mm9

Number of total reads
17275380
Reads aligned (%)
85.4
Duplicates removed (%)
18.5
Number of peaks
11167 (qval < 1E-05)

Base call quality data from DBCLS SRA