GSM2154777: bovine_SOX9_ChIPseq_1; Bos taurus; ChIP-Seq
Upon collection, gonads were crushed in liquid nitrogen using a mortar. Tissue powders were immediately fixed with PBS containing 2mM of Disuccinimidyl glutarate (DSG: PIERCE ref 20693) initially solubilized in DMSO as described previously (Nowak et al. 2005). Samples were then incubated for 30 minutes on a rotating wheel at room temperature. After three washes with PBS, samples were subsequently fixed with PBS 1% formaldehyde for 30 minutes at room temperature Libraries were prepared according to Illumina's instructions. 10-300ng DNA is combined with End Repair Mix, and incubate at 20°C for 30 min. Purify the end-repaired DNA with QIAquick PCR Purification Kit(Qiagen),then add A-Tailing Mix, incubate at 37°C for 30 min. Combine the purified Adenylate 3'Ends DNA, Adapter and Ligation Mix, incubate the ligation reaction at 20°C for 15 min. Purify the Adapter-ligated DNA with the QIAquick PCR Purification Kit. Several rounds of PCR amplification with PCR Primer Cocktail and PCR Master Mix are performed to enrich the Adapter-ligated DNA fragments. Then the PCR products are selected by running a 2% agarose gel to recover the target fragments. Purify the gel with QIAquick Gel Extraction kit (QIAGEN).
GEO Accession
GSM2154777