초록

Cell membranes and proteins are present in the PCR reaction solution due to the lack of a protein extractive step during the pretreatment. Nevertheless DNA amplification occurs under the presently claimed improved PCR method.

대표청구항 ▼

[ We claim:] [1.] A process for amplifying a specific nucleic acid sequence comprising the steps of:a) obtaining a blood sample, which includes a target deoxyribonucleic acid to be amplified;b) mixing said blood sample with plural primers, a polymerization agent, and mononucleotides, in a buffer sol

[ We claim:] [1.] A process for amplifying a specific nucleic acid sequence comprising the steps of:a) obtaining a blood sample, which includes a target deoxyribonucleic acid to be amplified;b) mixing said blood sample with plural primers, a polymerization agent, and mononucleotides, in a buffer solution with a pH between 8.1 to 9.5 as measured at 25.degree. C. to form a reaction mixture; wherein said oligonucleotide primers hybridize to opposite strands of the target deoxyribonucleic acid to be amplified;c) incubating said reaction mixture under conditions sufficient to release and denature any nucleic acids present;d) synthesizing an extension product of each primer, wherein the strands to which the primer was hybridized can serve as a template for the synthesis of the extension products thereby forming an extended hybrid;e) separating the extended hybrid into two single-stranded molecules by heat denaturation;f) treating the single-stranded molecules generated in step e) with the plural oligonucleotide primers and an excess amount of mononucleotides under conditions such that a second primer extension product is synthesized using each of the single-stranded molecules produced in step e) as a template; andg) repeating the steps e) and f) at least once, wherein said target deoxyribonucleic acid is amplified in the presence of impurities and in the absence of a protein extractive step.