Direct polymerase chain reaction (PCR) from human whole blood and filter-paper-dried blood by using a PCR buffer with a higher pH

被引:51
作者
Bu, Ying [1 ]
Huang, Huan [2 ]
Zhou, Guohua [1 ,3 ]
机构
[1] Huadong Res Inst Med & Biotechn, Nanjing 210002, Peoples R China
[2] China Pharmaceut Univ, Nanjing 210009, Peoples R China
[3] Nanjing Univ, Sch Med, Nanjing 210093, Peoples R China
基金
中国国家自然科学基金;
关键词
D O I
10.1016/j.ab.2008.01.010
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We described a novel approach to directly amplify genomic DNA from whole blood and dried blood spotted on filter paper without any DNA isolation by using the PCR buffer with a higher pH, which was optimized as pH 9.1-9.6. Direct PCR on blood treated with various anticoagulants showed that the buffer worked well with the blood treated by citrate, EDTA, or heparinate. DNA fragments with different lengths could be efficiently amplified directly from various forms of blood samples. By coupling the buffer with tetra-PCR, a '' true" single-tube genotyping was realized by using whole blood or paper-dried blood as starting material '' (c) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码:370 / 372
页数:3
相关论文
共 9 条
[1]  
Abu Al-Soud W, 2000, J CLIN MICROBIOL, V38, P345
[2]   Purification and characterization of PCR-inhibitory components in blood cells [J].
Abu al-Soud, W ;
Rådström, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (02) :485-493
[3]   Association of the Pro12Ala and C1431T variants of PPARG and their haplotypes with susceptibility to Type 2 diabetes [J].
Doney, ASF ;
Fischer, B ;
Cecil, JE ;
Boylan, K ;
McGuigan, FE ;
Ralston, SH ;
Morris, AD ;
Palmer, CNA .
DIABETOLOGIA, 2004, 47 (03) :555-558
[4]   Direct polymerase chain reaction from whole blood without DNA isolation [J].
Nishimura, N ;
Nakayama, T ;
Tonoike, H ;
Kojima, K ;
Kato, S .
ANNALS OF CLINICAL BIOCHEMISTRY, 2000, 37 :674-680
[5]  
Nishimura Naoyuki, 2002, Clin Lab, V48, P377
[6]   EFFECT OF HEPARIN ON POLYMERASE CHAIN-REACTION [J].
SATSANGI, J ;
JEWELL, D ;
WELSH, K ;
BUNCE, M ;
BELL, J .
LANCET, 1994, 343 (8911) :1509-1510
[7]   EFFECTS OF ANTICOAGULANTS AND STORAGE OF BLOOD-SAMPLES ON EFFICACY OF THE POLYMERASE CHAIN-REACTION ASSAY FOR HEPATITIS-C VIRUS [J].
WANG, JT ;
WANG, TH ;
SHEU, JC ;
LIN, SM ;
LIN, JT ;
CHEN, DS .
JOURNAL OF CLINICAL MICROBIOLOGY, 1992, 30 (03) :750-753
[8]   Inhibition and facilitation of nucleic acid amplification [J].
Wilson, IG .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1997, 63 (10) :3741-3751
[9]   An efficient procedure for genotyping single nucleotide polymorphisms [J].
Ye, S ;
Dhillon, S ;
Ke, XY ;
Collins, AR ;
Day, INM .
NUCLEIC ACIDS RESEARCH, 2001, 29 (17) :88-88