Methylation profiling by high throughput sequencing
Summary
Deep BS-Seq of two Ants: Camponotus floridanus and Harpegnathos saltator.
Overall design
Up to 20 µg of total genomic DNA were isolated from embryos, larvae, males, virgin queens and queens for both species, as well as major and minor workers for C. floridanus, and workers and gamergates for H. saltator. DNA was fragmented by sonication with a Covaris S2 system (Covaris, MA) to a mean size of approximately 150 bp, followed by end-repair, 3'-end addition of dA, and adapter ligation. Methylated adapters were used according to the manufacturer's instructions (Illumina). The bisulfite conversion of sample DNA was carried out using a modified NH4SO4-based protocol and amplified by 9 cycles of PCR. Pair-end sequencing was carried out using Illumina Genome Analyzers.
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