ChIP-Atlas: SRX1650677

Antigen

Cell type Class: Epidermis
Cell type: Keratinocytes
MeSH Description: Epidermal cells which synthesize keratin and undergo characteristic changes as they move upward from the basal layers of the epidermis to the cornified (horny) layer of the skin. Successive stages of differentiation of the keratinocytes forming the epidermal layers are basal cell, spinous or prickle cell, and the granular cell.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. Chromatin immunoprecipitated DNA was prepared for sequencing following a modified version of Illumina's Genomic DNA protocol. Sheared DNA fragments undergo end repair by T4 DNA polymerase to fill in 5′ overhangs, Klenow polymerase to remove 3′ overhangs, and T4PNK to phosphorylate the 5′-OH. For Illumina adapter ligation, a single adenine nucleotide overhang must be added to the polished DNA. A 1:30 dilution of the Adaptor Oligo Mix (20) was used in the ligation step. The resulting adapted constructs are amplified by 18 cycles of PCR enriching the DNA fragments with adapters flanking each end. The amplified product was quantified using a spectrophotometer at 260 nm (Nanodrop) then a portion of final sample is size separated on a 6% TBE (Invitrogen) to verify correct target range amplification. The validated DNA library is submitted at a 10 nmol/L concentration for sequencing. All protocols for Illumina library sequence preparation, sequencing, and quality control are provided by Illumina (20).