Direct polymerase chain reaction (PCR) from human whole blood and filter-paper-dried blood by using a PCR buffer with a higher pH.
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.
Bu
Ying
Y
Huadong Research Institute for Medicine and Biotechnics, Nanjing, 210002, China.
Huang
Huan
H
Zhou
Guohua
G
D001769:Blood / Q000378:metabolism*; D002021:Buffers; D004247:DNA / Q000235:genetics; D046209:Electrophoresis, Microchip; D015894:Genome, Human / Q000235:genetics; D005838:Genotype; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008851:Micropore Filters; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D002021:Buffers; D004247:DNA