GSM2536473: b-catenin mutant cranial mesecnhyme + ectoderm; Mus musculus; ChIP-Seq
At E13.5, En1Cre;b-cateninfl/fl mutant and En1Cre;b-cateninfl/+ control cranial mesenchyme was isolated by manual dissection. An incision was made around the circumference of the neurocranium, and the tissue covering the brain was manually disassociated. The supraorbital cranial mesenchyme isolated consists of the neural crest and mesoderm derived cranial mesenchyme plus the ectoderm. 14 µg chromatin was immunoprecipitated with 4 µg rabbit anti-H3K27me3 (Millipore #07-449). Illumina sequencing libraries were prepared from the ChIP and Input DNAs by the standard consecutive enzymatic steps of end-polishing, dA-addition, and adaptor ligation. After a final PCR amplification step, the resulting DNA libraries were quantified and sequenced on NextSeq 500. The libraries were made using the standard Illumina PE adaptor: ACACTCTTTCCCTACACGACGCTCTTCCGATC*T; p-GATCGGAAGAGCGGTTCAGCAGGAATGCCGAG