Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Unclassified
Antigen
Unclassified

Cell type

Cell type Class
Muscle
Cell type
Myotube
NA
NA

Attributes by original data submitter

Sample

source_name
Myotubes
sample type
Myotubes

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Chromatin immnunoprecipitation sequencing (ChIP-Seq) was performed on 20 mg cell lysate using the following antibodies: anti H3K4me3, Cat # 8580 Abcam; anti H3K27me3, Cat # 07-449, Millipore; anti H3K4me1, Cat # 5326, Cell Signaling; anti histone H3, Cat # 05-499, Millipore. ChIP library was constructed as described previously (Soleimani, V.D., et al., Transcriptional dominance of Pax7 in adult myogenesis is due to high-affinity recognition of homeodomain motifs. Dev Cell, 2012. 22(6): p. 1208-20.). Sequencing was done on Illumina GAIIX Genome Analyzer in duplicates. ChIP-Seq of Sox2 was performed by Chromatin Tandem Affinity Purification Sequencing as described previously (Soleimani, V.D., et al., Transcriptional dominance of Pax7 in adult myogenesis is due to high-affinity recognition of homeodomain motifs. Dev Cell, 2012. 22(6): p. 1208-20. Soleimani, V.D., et al., Chromatin tandem affinity purification sequencing. Nat Protoc, 2013. 8(8): p. 1525-34) . Briefly, to produce the TAP-tagged fusion construct mouse Sox2 gene was fused with a C-terminal TAP tag (6XHIS-TEV-3FLAG) and inserted into Sox2 locus using homologous recombination in G4 ES cell line (G4-129S6B6F1, Samuel Lunenfeld Research Institute, Mount Sinai Hospital). ES cells harboring the recombinant knocked-in allele of Sox2 was grown on Mitomycin C treated MEFs in the presence of LIF. RNA-Seq: Total RNA was extracted from primary myoblasts or differentiated myotubes over expressing and empty vector retroviral construct (48 hours differentiation) as described previously (Soleimani, V.D., et al., Snail regulates MyoD binding-site occupancy to direct enhancer switching and differentiation-specific transcription in myogenesis. Mol Cell, 2012. 47(3): p. 457-68.).

Sequencing Platform

instrument_model
Illumina Genome Analyzer IIx

mm10

Number of total reads
76908584
Reads aligned (%)
97.5
Duplicates removed (%)
13.5
Number of peaks
792 (qval < 1E-05)

mm9

Number of total reads
76908584
Reads aligned (%)
97.3
Duplicates removed (%)
13.5
Number of peaks
937 (qval < 1E-05)

Base call quality data from DBCLS SRA