title: GSM2836707: ChIPseq amphioxus H3K4me3 15h replicate 2; Branchiostoma lanceolatum; ChIP-Seq
library_construction_protocol: Batches of embryos from six dishes were transferred into a 2mL tube. After a quick centrifugation at 9000 rpm, the seawater was removed and replaced by 1.5mL of fixation solution (1.85% formaldehyde in MOPS-EGTA buffer: 0.1M MOPS pH 7.5, 2mM MgSO4, 1 mM EGTA, 0.5M NaCl). After 15 minutes at room temperature, 155 µL of Glycine 10% was added and the embryos were allowed to naturally sediment. The solution was then gently removed and the embryos were washed five times at 4°C with cold NaPBS (0.9% NaCl, 20 mM sodium phosphate buffer at pH 7.4). After a quick centrifugation at 13,000 rpm, the liquid was removed and the fixed embryos were frozen in liquid nitrogen and subsequently stocked at -80°C. For the chromatin immunoprecipitation, embryos were homogenized and lysed in order to release the chromatin, which was subsequently sonicated in a Bioruptor (Diagenode). Then the sonicated product was incubated with the specific antibody first, and then with Dynabeads (Thermofisher, cat. 10007D). After washing, pulled down chromatin was eluted from the beads and decrosslinked. Illumina libraries were prepared following manufacturer's instructions.
attributes.tag: GEO Accession
attributes.value: GSM2836707