The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.
The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.
Patent No.:
Date of Patent:
Aug. 10, 1999
Filed:
Nov. 18, 1994
Naoyuki Nishimura, Kyoto, JP;
Tomoko Nakayama, Osaka, JP;
Shimadzu Corporation, Kyoto, JP;
Abstract
This invention is directed to a novel method for PCR amplification wherein PCR is carried out at a higher pH than the pH widely used in the art. Specifically, the buffer solution is adjusted to pH 9.0 to 11.0 at 25.degree. C. Using the present invention, DNA amplification can be successfully carried out following a simple pretreatment. In the present invention whole blood is mixed with a hypotonic solution so that a selective lysis of red blood cells takes place. The residual leukocytes are then collected. The leukocytes are mixed with a polymerization agent, primers and other necessary reagents and PCR is carried out. When the PCR solution is placed at a high temperature for DNA denaturation, the leukocytes are lysed so that the leukocyte DNA is released and can access the primers and the other necessary reactions for PCR in the solution. 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.