Abstract
The isolation of high quality genomic DNA (gDNA) is the precursor to many molecular biology assays. The new GenFind V3 Blood and Serum DNA Isolation Kit from Beckman Coulter uses the patented SPRI (Solid Phase Reverse Immobilization) paramagnetic bead technology to isolate genomic DNA from fresh or frozen whole blood and serum containing Citrate, EDTA, or Heparin anticoagulants, as well as from cultured cells. GenFind V3 uses an improved cell lysis buffer and Proteinase K treatment to rupture cell membranes and digest proteins to give consistently high quality gDNA with improved yields and purities. The GenFind V3 kit supports up to 400L sample input volumes from blood or serum or two million cultured cells and can be performed in either a 96-well plate or tube-based format. Additionally, the need for large volume whole blood extractions continues to grow along with the ability to easily automate the process to alleviate user errors. To address this need, a protocol is available that supports >400L and up to 2 mL of whole blood is available that can be performed in an automated fashion using a 24-well plate. Volumes greater than 2 mL can be performed in a tube-based format.
Here, we demonstrate a walk-away automated solution for GenFind V3 kit to purify up to 400L of whole blood using the Beckman Coulter Biomek i7 Hybrid liquid handling system. The Biomek i-Series method is a high yielding and robust nucleic acid purification process and can process up to 96 samples in a 96-well format in less than 3.5 hours with minimal user interaction and no off-deck centrifugation or vacuum filtration. The method is also compatible with the Biomek i-Series NGS workstation, facilitating the installation process for existing Biomek i-Series users.