GSM1815460: Input DNA GAF; Drosophila melanogaster; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Larvae
Cell type
Salivary glands
NA
NA
Attributes by original data submitter
Sample
source_name
Or-R
genotype
wildtype
tissue
Imaginal discs and salivary glands
developmental stage
3rd instar stage
antibody
none (input)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Lysates were prepared from sonicated nuclei and GAF-DNA complexes were isolated with antibody. The GAF antibodies were custom-made by this lab while anti-RNA Pol II antibodies were provided by the lab of Dr. John T. Lis. ChIP-seq libraries were prepared using the Illumina ChIP-seq Library preparation Kit following the manufacture’s instructions with minor modifications. Briefly 10ng of purified ChIP DNA was end repaired by conversion of overhangs to phosphorylated blunt ends. A’ bases were added to the 3’ends of the DNA fragments and Illumina adapters (1:30 dilution) were ligated to the ends of ChIP fragments. This fragment range corresponds to a ChIP fragment range of about 200 -300bp. Size selected fragments were PCR amplified for 12 cycles . Libraries were quantified and validated using Agilent Technologies 2100 Bioanalyser for size, concentration and purity. Q-PCR was repeated to confirm retention of relative enrichment.