Drosophila miranda MSH22 female genomic DNA Illumina paired-end library
We extracted genomic DNA from 50 pooled male flies or female flies, and library preparation followed the manufacturer's instructions (Illumina). Briefly, 2–5 µg of genomic DNA diluted in 50 µl TE buffer were fragmented by nebulization, generating double-stranded DNA fragments with blunt ends or with 3' or 5' overhangs. The overhangs were converted to blunt ends using T4 DNA polymerase and Klenow polymerase, after which an ‘A’ base was added to the ends of double-stranded DNA using Klenow exo– (3' to 5' exo minus). Next, DNA adaptors (Illumina) with a single ‘T’ base overhang at the 3' end were ligated to the above products. These products were then separated on a 2% agarose gel, excised from the gel at a position of around 200bp, and purified (Qiagen Gel Extraction Kit). The adaptor-modified DNA fragments were enriched by PCR with PCR primers 1.1 and 2.1 (Illumina). The concentration of the libraries was measured by absorbance at 260 nm and the fragment size invesitgated on Agilent 2100 Bioanalyzer.