DNA Extraction

Prior to extraction, samples should be randomized, and each batch should include five extraction blanks.

We use two kit-based DNA extraction methods depending on sample throughput. Prior to sample homogenization, all samples undergo a 10-minute incubation at 65 °C. All other steps follow the manufacturer's instructions unless otherwise noted.

For small batches (up to ~24 samples), we use the DNeasy® PowerSoil® Pro Kit (Qiagen, Hilden, Germany; Cat. Nos. 47014 and 47016).

For larger batches (up to 96 samples), we use the MagAttract® PowerSoil® Pro DNA EP Kit (384) (Qiagen, Hilden, Germany; Cat. No. 47119) in combination with an epMotion 5075 (Eppendorf, Hamburg, Germany). We typically extract no more than 88 samples per run, leaving the 12th column of a 96‑well plate intentionally empty to accommodate PCR controls. These extractions are usually completed over two days by incubating the lysate with Solution CD2 at 4 °C overnight.

Note

Some sample storage solutions may be incompatible with these extraction kits.

With extraction complete, proceed to PCR Amplification.