Puregene tissue dna extraction kit

DNA from plastics was extracted using a modified bead-beating approach combined with the Puregene tissue DNA extraction kit (Qiagen, Valencia, CA) as previously described (69 (link)). Samples with a sufficient amount of DNA were single-end sequenced (1 × 150 bp) with Illumina NextSeq technology at Microsynth AG, Switzerland. These were plastic A right after collection (A0), after 1 year of incubation (A1), and after 2 years of incubation (A2), plastic C after 1 year and 2 years of incubation (C1 and C2, respectively), and plastic B after 1 year of incubation (B1). All raw sequences files are available from the NCBI Sequence Read Archive (SRA) database (BioProject PRJNA599974). Due to low DNA concentrations, samples from plastics B and C right after collection (samples B0 and C0) and sample B2 were not sequenced.

From each pool of 100 embryos, DNA was purified with the Puregene tissue DNA extraction kit (Qiagen, Germany), with modifications described in SI materials and methods. DNA was assessed for RNA contamination and integrity with gel electrophoreses and measured for concentration and quality by Nanodrop (ND-1000; Thermo Scientific). DNA was stored at −20 °C until further analysis.