GSM3097754: R41_NA_N_120min_h3k27ac; Apis mellifera; ChIP-Seq
Bee heads were separated from bodies on dry ice and placed in a dissection dish with pure ethanol and dry ice to prevent degradation during dissection. The cuticle of the head capsule was removed and each head was placed in PBS + Protease Inhibitor Cocktail (Roche) at -20ºC for 14 to 18 h. The heads were then opened, the hypopharyngeal glands removed, and the whole brain removed.  The brains were placed in a new 1.5ml tube and kept frozen (-80°C) until ChIP. ChIP was performed with a Diagenode iDeal ChIP kit and the standard protocol using Abcam ab4729 (rb polyclonal anti-H3K27Ac) antibodies. 1/10 volume input  DNA samples were collected from each aliquot and pooled and used as a background control for peak calling ChIP-Seq libraries were constructed with KAPA LTP library kits (KK8230) The libraries were pooled in equimolar concentration as per instructions and each pool was quantitated by qubit (invitrogen). Single-end sequencing (read length = 100nt) was performed on an Illumina HiSeq2000 using a TruSeq SBS sequencing kit version 3.
GEO Accession
GSM3097754