FAQ: Can the Monarch HMW DNA Extraction Kits be used to process human Buffy coat?

Yes, the Monarch HMW DNA Extraction Kit for Cells & Blood (NEB #T3050) can be used for processing Buffy coat. When working with Buffy coat, which is a concentrated PBMC solution, smaller input amounts should be used than when working with blood. The total cell count should not be higher than 1 x 107 cells for the standard protocol (agitation at 2000 rpm) and not higher than 5 x 106 cells when UHMW DNA is needed (low agitation). Buffy coat as obtained from blood banks contains up to 1 x 109 PBMCs per 30-80 ml, which equates to approximately 1 x 107 -3 x 107  cells/ml. We suggest starting with 100-300 μl of this solution, add PBS to a total volume of
500 μl, and move into the erythrocyte lysis procedure as if it were a 500 μl blood sample.
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    Tips for HMW DNA Extraction from Blood Samples with the Monarch® kit (NEB #T3050)

     In this video, we share some important tips for successfully extracting high molecular weight DNA (HMW) from blood samples.