A Simple and Efficient Method for DNA Purification from Samples of Highly Clotted Blood |
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Authors: | Ruyi Xu Ping Ye Leiming Luo Hongmei Wu Jin Dong Xinxin Deng |
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Institution: | (1) Department of Geriatric Cardiology, Chinese PLA General Hospital, 28 Fuxing Rd, Beijng, 100853, China;(2) Department of Biochemistry, Chinese PLA General Hospital, Beijing, China; |
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Abstract: | Rapid purification of DNA from samples of highly clotted blood is a challenging problem due to the difficulty in recovering
and dispersing blood clots. We developed a new method for discarding the serum-separator gel and rapidly dispersing the blood
clots. A special disposable tip was inserted into the serum-separator gel so that the serum-separator gel could be discarded.
The blood clot obtained was dispersed into small pieces through a copper mesh (pore size, 250 μm) in a special dispersing
instrument by centrifugation. After lysis of red blood cells and white blood cells, genomic DNA was concentrated and desalted
by isopropanol precipitation. The mean yield of DNA purified from a 0.3-ml blood clot was 22.70 μg in 173 samples of clotted
blood cryopreserved for 1 month, and 19.02 μg in 1,372 samples of clotted blood cryopreserved for >6 months. DNA samples were
successfully performed through polymerase chain reaction, real time polymerase chain reaction, and melt curve analysis. Their
quality was comparable with that purified directly from EDTA-anticoagulated blood. The new method overcomes the difficulties
in recovering and dispersing blood clots, allowing efficient purification of DNA from samples of highly clotted blood. |
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