ChIP-Atlas: SRX3330450

Antigen

Cell type Class: Epidermis
Cell type: Foreskin
MeSH Description: The double-layered skin fold that covers the GLANS PENIS, the head of the penis.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: ChIP was performed with the SimpleChIP Enzymatic Chromatin IP Kit w/ Magnetic Beads (Cell Signaling). Protein-DNA complex were crosslinked, DNA was digested with MNase to yield 150-900bp fragments and Myc-UL34 bound DNA was isolated with antibody then purified. Libraries were constructed with the TruSeq ChIP Sample Preparation Kit (Illumina). Briefly, ChIP-enriched DNA were fragmented with Covaris Ultrasonicator and fragment sizes were confirmed on Agilent High Sensitivity DNA chip on an Agilent 2100 Bioanalyzer. DNA ends were repaired to generate dsDNA moleculars and purified with Agencourt AMPure XP beads. The repaired dsDNA were adenylated and adaptors for Illumina sequencing were added to the A-overhang prior to repeat purification with Agencourt AMPure XP beads. Bead-purified samples were size selected on Pippen Prep targeting 150-350 bp. The ligated library was enriched with 18 cycles of PCR and purified again with Agencourt AMPure XP beads. Size distribution was confirmed again with Agilent Bioanalyzer; the exact quantification was performed with Kapa Biosystem Complete Kit (ROX Low) on a Stratagene Mx3000P Real Time PCR machine. Enriched libraries were normalized and pooled in equimolar ratios, denatured and poolde with PhiX control (10%) prior to sequencing on an Illumina MiSeq with 2 x 75 Paired End v3 chemistry.