GSM8486598: Cp, H3K27ac, developing distal hindlimb, wt, CS19, Rep1; Carollia perspicillata; ChIP-Seq
Mouse and bat embryonic limbs were dissected and fixed in 1% formaldehyde in 10% FCS/PBS and subsequently snap-frozen and stored at −80°C until further processing Chromatin immunoprecipitations were performed using the iDeal ChIP-seq Kit for Histones (Diagenode, #C01010051) and iDeal ChIP-seq Kit for Transcription Factors (Diagenode, #C01010055) according to the manufacturer's instructions. Briefly, fixed limbs were lysed and sonicated using a Bioruptor Plus Sonication device (45 cycles, 30s on, 30s off, at high power setting) in provided buffers. 5 µg sheared chromatin were used for histone immunoprecipitation with 1µg of anti-H3K27ac (Diagenode, #C15410174). For Meis2 immunoprecipitation, 20 µg of sheared chromatin were used with two anti-Meis antibodies simultaneously, one recognizing the conserved C-terminal domain of Meis1a and Meis2a and the other recognizing all Meis2 isoforms as previously described26. 2µg of each antibody were used per immunoprecipitation. Antibodies were generously provided by Dr. Miguel Torres. Libraries were prepared using the Kapa HyperPrep Kit (Roche, #07962347001) and libraries were sequenced on a NovaSeq2 with 100bp paired-end reads. ChIP-seq experiments were performed in biological duplicates.