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 (one or more 96‑well plates), 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 83 samples per run to accommodate extraction blanks and 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.