Document Detail


A simple and efficient method for DNA purification from samples of highly clotted blood.
MedLine Citation:
PMID:  20549389     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Ruyi Xu; Ping Ye; Leiming Luo; Hongmei Wu; Jin Dong; Xinxin Deng
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Molecular biotechnology     Volume:  46     ISSN:  1559-0305     ISO Abbreviation:  Mol. Biotechnol.     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-09-29     Completed Date:  2011-02-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9423533     Medline TA:  Mol Biotechnol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  258-64     Citation Subset:  IM    
Affiliation:
Department of Geriatric Cardiology, Chinese PLA General Hospital, 28 Fuxing Rd, Beijng, 100853, China.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Blood Coagulation*
DNA / blood,  isolation & purification*
DNA Primers
Electrophoresis, Agar Gel
Humans
Polymerase Chain Reaction
Chemical
Reg. No./Substance:
0/DNA Primers; 9007-49-2/DNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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