A key source for genomic DNA(gDNA) is blood drawn into a standard 10mL Vacutainer® tube. The Promega ReliaPrep™ Large Volume HT gDNA Isolation Syste integrated on the Hamilton MICROLAB® STARplus liquid handling workstation provides a unique and dependable system for isolating genomic DNA from large volumes (3 mL–10 mL) of blood. The novel chemistry and instrumentation resolve many challenges encountered when processing large-volume samples in a high-throughput format such as: loss of sample pellets during decanting of fluids, transport of full 50 mL tubes to various locations on a liquid handling robot, and manual re-suspension of final DNA pellets. Liquid handler resource constraints were removed by creation of a new accessory, the ReliaPrep HSM 32 LV instrument, which provides heating, shaking and magnetization of samples at one deck position. The combination of this device, the MICROLAB STARplus workstation and the ReliaPrep Large Volume HT gDNA Isolation System allows automated recovery of pure gDNA from up to 96 ten milliliter blood samples within 8 hours. We present verification studies demonstrating automated system performance. Comparisons between the ReliaPrep™ Large Volume HT gDNA Isolation System and a standard precipitation-based method were made for duplicate blood samples from multiple donors. Yield, purity, and integrity of extracted gDNAwere assessed using UV absorbance spectroscopy and gel electrophoresis. Genomic DNA yields from normal 10 mL whole blood samples were 200 400 μg (depending on white blood cell count) in an eluted volume of 1mL. Recovered DNA exhibited good purity with A260/A280 ratios greater than 1.7 and A260/A230 ratios between 1.8 and 2.2. Isolated DNA was suitable for storage and was used in many downstream analysis applications. Results of genomic DNA purification from frozen (hemolysed) blood samples and blood collected using common anticoagulants (EDTA, heparin, citrate)are also compared to demonstrate the efficacy of the new system.
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