title: GSM6208352: E14.5 mouse kidney, IgG, biol rep 2; Mus musculus; ChIP-Seq
library_construction_protocol: E14.5 kidneys were harvested and fixed with 2% formaldehyde for 10 minutes at room temperature. After quenching and cell lysis by Dounce homogenizer, chromatin was sonicated using a Covaris S220 focused-ultrasonicator for 7.5 minutes at 7.5 Watts, peak power 75 and duty factor 10. ChIP-seq libraries were prepared using the KAPA Hyperprep Kit using the xGen Stubby Adaptor and Illumina Index Primers. Libraries amplification cycle number was determined by performing a test amplification on 5% of the volume of each library for 40 cycles. The final amplification cycle number was the threshold cycle number +3 cycles (16 cycles for input samples, 20 cycles for IgG and HNF-1β IP samples). Post amplification libraries were size-selected at 200-400bp in length by gel electrophoresis.