Automated DNA Extraction from Cells

Semi-automated DNA extraction was performed on a Gilson PIPETMAX liquid handling robot enabled for exclusion-based sample preparation (ESP), termed EXTRACTMAX [50 (link)]. LiDS buffer (90 mM Tris–HCL, 500 mM lithium chloride, 1% Igepal CA-630, 10 mM EDTA, 1 mM dithiothreitol) and MagneSil Paramagnetic Particles (PMPs) (Promega) resuspended in GTC buffer (10 mM Tris–HCl, 6 M guanidinium thiocyanate, 0.1% Igepal CA-630, pH 7.5) are added to the EXTRACTMAX extraction microplate (Gilson) by the robot. Cells were added to the microplate well containing LiDS, GTC, and MagneSil beads and mixed by the robot. Cells were allowed to lyse, and DNA was allowed to bind to MagneSil PMPs for 5 min. The robot then transferred the MagneSil PMPs with bound DNA by exclusion liquid repellency (ELR) through one PBST (PBS containing 0.1% Tween-20) wash, one PBS wash, and into water for elution. Beads were manually resuspended in the elution well and allowed to elute for 2 min. The MagneSil PMPs are magnetically transferred out of the elution well, leaving eluted DNA in water. LNCaP and WBC DNA for restriction enzyme and pre-amplification validation experiments were extracted using the AllPrep DNA/RNA mini kit (Qiagen) according to manufacturer’s instructions.