GSM2813259: A1_5min; Apis mellifera ligustica; Bisulfite-Seq
Heads were partially dissected to expose brain and submerged in RNALater ICE overnight at -20°C. Whole brains were dissected the following day and stored at -80C until DNA extraction. Genomic DNA was extracted from whole brains using the Gentra Puregene Tissue Kit (Qiagen) and stored at -20°C.  Unmethylated lambda phage DNA (Promega) was spiked in as a control for assessment of bisulfite conversion efficiency. The samples were submitted to the Roy J. Carver Biotechnology Center at the University of Illinois for quality control, library construction, and sequencing.  The integrity of the samples was checked using the AATI Fragment Analyzer, which confirmed that the DNA was of high molecular weight.  Shotgun DNA libraries were made using the KAPA Library Preparation Kit, then bisulfite treated using the Zymo EZ DNA Methylation-Lightning Kit.  The libraries were then amplified for 10 PCR cycles using HiFi Uracil+ DNA Polymerase (KAPA Biosystems), pooled in equimolar quantities, and sequenced on the HiSeq 2500 (Illumina).  Paired-end 100-nucleotide sequence reads were generated from seven control and seven experimental samples for each collection time point (5 minutes and 120 minutes).
GEO Accession
GSM2813259