A simple and efficient method for DNA purification from samples of highly clotted blood.
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.
Xu
Ruyi
R
Department of Geriatric Cardiology, Chinese PLA General Hospital, 28 Fuxing Rd, Beijng, 100853, China.
Ye
Ping
P
Luo
Leiming
L
Wu
Hongmei
H
Dong
Jin
J
Deng
Xinxin
X
D001483:Base Sequence; D001777:Blood Coagulation; D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D006801:Humans; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004247:DNA