title: GSM1150348: FaireSeq_NT_2; Mus musculus; OTHER
library_construction_protocol: Tissue samples from E12.5 neural tube and E17.5 pancreas were dissected in cold PBS, homogenized, and crosslinked with 1% formaldehyde for 8 minutes at room temperature and fixation was terminated by adding glycine to a final concentration of 0.125 M. Samples were sonicated using a Diagenode bath Sonicator for 36 minutes, with 30 second on:off cycle times. Samples were centrifuged at 14,000 rpm to pellet debris. Two rounds of phenol:chloroform extraction were performed yielding 500 μl aqueous phase, to which 8.25 μl 5M NaCl solution was added. Samples were incubated overnight at 65 oC to reverse crosslinks. Samples were ethanol precipitated and resuspended in 50 μl 10mM Tris-HCl pH 7.5, digested for 4 hours using 5 μl 10 mg/ml Proteinase K. 1 μl of 10 mg/ml RNAse A was added, and incubated an additional 30 minutes at 37 oC. Samples were purified using a Qiagen PCR Cleanup kit. All libraries were made according to NEBNext ChIP-Seq Library Prep Set Master Mix for Illumina protocol