ena-STUDY-The Pirbright Institute-06-06-2021-18:44:07:101-706
Analysis of the open chromatin regions in early embryos of the African malaria mosquito Anopheles gambiae
Anopheles gambiae were maintained at 28 °C and 80% humidity. Females were blood-fed and three days later allowed to lay eggs for two hours. After oviposition and incubation for additional two hours in the insectary conditions, the embryos were washed and crosslinked using formaldehyde. The nuclei were released using Dounce homogeniser, pelleted and, following resuspension in FAIRE Lysis Buffer, the chromatin was released by bead beating. Chromatin was then sonicated using Fisherbrand Q500 Sonicator to generate approximately 300-400 long fragments. The phenol-chloroform isolated DNA fraction was used to generate a library sequenced on an Illumina NovaSeq6000 platform to obtain approximately 150 million 150 bp long paired-end reads for each of three replicates.
Anopheles gambiae were maintained at 28 °C and 80% humidity. Females were blood-fed and three days later allowed to lay eggs for two hours. After oviposition and incubation for additional two hours in the insectary conditions, the embryos were washed and crosslinked using formaldehyde. The nuclei were released using Dounce homogeniser, pelleted and, following resuspension in FAIRE Lysis Buffer, the chromatin was released by bead beating. Chromatin was then sonicated using Fisherbrand Q500 Sonicator to generate approximately 300-400 long fragments. The phenol-chloroform isolated DNA fraction was used to generate a library sequenced on an Illumina NovaSeq6000 platform to obtain approximately 150 million 150 bp long paired-end reads for each of three replicates.