GSM4905020: Homozygous dpolh mutant_part1; Drosophila melanogaster; OTHER
Each fly was crushed with a pestle in a 1,5 mL-1 tube containing 170 µL of extraction buffer (Tris-HCl pH 8.0; 50mM; EDTA 50mM; SDS 1 %). Proteinase K (555 µg µL-1) was added once the tissues had been completely grinded. After incubating 15 min at room temperature, cellular debris was removed twice by potassium acetate addition (0,83 M) and centrifugation with a bench-top centrifuge (max speed) at 4 °C for 10 min. DNA was isolated by double phenol/chloroform/isoamyl alcohol (25:24:1) extraction and ethanol precipitation overnight at -20 °C with glycogen (20 µg) followed by centrifugation at 4 °C. Precipitated DNA was washed with cold ethanol 70 %, dried at room temperature for 30 min and dissolved in water. The quality of extracted DNA (about 150 ng) was verified by agarose gel electrophoresis. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit