Patent ID: 6673578
Filing Date: 2004-01-06
Classification: C12N,C12Q

Abstract:
A DNA synthesis method with a shortened time period for DNA synthesis by polymerase chain reaction (PCR), comprising the steps of:carrying out a DNA synthesis by PCR with a reaction mixture comprising: DNA polymerase, template DNA, dNTPs, and primers, wherein the amount of DNA polymerase added to said reaction mixture is pre-determined for each specific type of DNA polymerase by carrying out a test PCR reaction under the following conditions (A) and (B): (A) reaction mixture: 50 &mgr;l volume of a reaction mixture comprising DNA polymerase, 1 ng of genomic DNA from Escherichia coli, and 10 pmol each of primers Eco-1 (SEQ ID NO: 10) and Eco-2 (SEQ ID NO: 11); and having a composition suitable for said DNA polymerase; and (B) reaction conditions: 35 cycles of PCR, wherein one cycle consists of 99Â° C., 1 second-66Â° C., 7 seconds; wherein the amount of DNA polymerase used is that which is sufficient to provide more than 10 ng of amplified DNA fragments of about 2 kb per 50 &mgr;l of reaction mixture.