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Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood ,,, Mini Kit

Laura Pollock, Jennifer Halcome, and Michele Nelson Hurst
Eppendorf 5 Prime, Inc. Abstract

The Perfect gDNA Blood Mini Kit is an efficient way to purify genomic DNA from 200 l of whole blood as described in the product manual. In addition, it has been shown that the kit can also be used to effectively purify genomic DNA from buffy coat after the whole blood has been separated into its components. Using this method provides obvious benefits to the researcher because the gDNA isolated from buffy coat is higher in concentration and yield than the corresponding whole blood sample. Best results are found when an entire vial of blood is separated into its component parts, but it has also been proven that the buffy coat, from as little as 1.5 ml of whole blood, gives good results.

Introduction

The versatile Perfect gDNA Blood Mini Kit allows the researcher to isolate gDNA from a variety of biological samples, including buffy coat. The purification method is based on Proteinase K lysis followed by collection of the crude high molecular weight gDNA on a membrane. The gDNA is then washed to remove contaminants and eluted in a buffer suitable for common downstream applications such as PCR and restriction digest. The method is spin-column-based and simply requires the use of a standard microcentrifuge. No additional purification is required.

Buffy coat is the component of whole blood that contains leukocytes and platelets and makes up only 1% of the entire whole blood composition. Separating out this particular component enables the researcher to enrich the DNA-containing leukocytes and
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