DNA Extraction from small blood volumes and the processing of cellulose blood cards for use in polymerase chain reaction |
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Authors: | Karol Mackey Amy Steinkamp Piotr Chomczynski |
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Affiliation: | (1) Molecular Research Center, Inc., 5645 Montgomery Road, 45212 Cincinnati, OH;(2) Division of Endocrinology, College of Medicine, University of Cincinnati, Cincinnati, OH |
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Abstract: | This article describes two procedures for the purification of genomic DNA from small blood volumes of whole blood using DNAzol®BD. In the first procedure, DNA is isolated from 1–20 μL of whole blood using a fast and simple protocol that is appropriate for the simultaneous extraction of a large number of samples. The isolated DNA is suitable for gel electrophoresis and polymerase chain reaction (PCR). In the second procedure, cellulose blood cards containing approx 5 μL of dried blood are treated with DNAzol BD in order to retain DNA on the cellulose matrix while removing other cellular components. The blood card with DNA subsequently serves as template in PCR. The blood card processing and amplification procedures are performed in the same PCR tube without any centrifugation steps, making the combined procedures amenable for automated DNA preparation and amplification in a single tube. |
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Keywords: | Blood blood cards DNA DNA extraction polymerase chain reaction PCR small blood volume Guthrie cards |
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