Zymo quick dna fecal soil microbe kit

DNA extraction was completed using the Zymo Quick-DNA Fecal/Soil Microbe Kit (Zymo Research Corporation, Irvine, CA, USA) with the following modifications. First, samples were placed in a 2 ml MP Biomedicals (Santa Ana, CA, USA) Lysing Matrix A tube with 750 µl of BashingBead™ buffer and then placed in a Bead Ruptor 12 (Omni International, Inc., Kennesaw, GA, USA) for two 90-s rounds on high. Final elution was performed using 75 µl of 10 mM Tris–HCl buffer (pH 8.5; bioWORLD, Dublin, OH, USA). DNA quantification was performed at the University of Kentucky Genomics Core Laboratory using the Qubit 2.0 (Thermo Fisher Scientific, Waltham, MA, USA).

Three DNA extraction kits were tested on eggs and testes using the manufacturer’s instructions. The prepGEM Insect DNA extraction kit (ZyGEM Corporation Ltd, Hamilton, New Zealand) was used on 14 male samples and two female samples, the Zymo Quick DNA Fecal/Soil Microbe kit (Zymo Research, California, USA) was used on three male samples and the NucleoSpin DNA purification kit (Macherey–Nagel, Düren, Germany) was used on 13 female samples.