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    • 48. 发明授权
    • Method for synthesizing DNA
    • DNA合成方法
    • US06673578B1
    • 2004-01-06
    • US09786684
    • 2001-05-22
    • Takashi UemoriYoshimi SatoMariko OkawaTomoko FujitaKazue MiyakeOsamu TakedaHiroaki SagawaMichio HagiyaHiroyuki MukaiKiyozo AsadaIkunoshin Kato
    • Takashi UemoriYoshimi SatoMariko OkawaTomoko FujitaKazue MiyakeOsamu TakedaHiroaki SagawaMichio HagiyaHiroyuki MukaiKiyozo AsadaIkunoshin Kato
    • C12P1934
    • C12N15/1003C12Q1/686C12Q2527/125
    • A DNA synthesis method with a shortened time period required for DNA synthesis by polymerase chain reaction (PCR), characterized in that a DNA polymerase is used in an amount effective for providing more than 10 ng of amplified DNA fragments of about 2 kb per 50 &mgr;l of a reaction mixture, when PCR is carried out 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 and Eco-2 (nucleotide sequences of the primers Eco-1 and Eco-2 being shown in SEQ ID NOs: 10 and 11 of Sequence Listing, respectively); and having a composition suitable for the DNA polymerase; and (B) reaction conditions: 35 cycles of PCR, wherein one cycle consists of 99° C., 1 second-66° C., 7 seconds; a kit for DNA synthesis usable for the DNA synthesis method; and an article of manufacture of a PCR agent. According to the present invention, the procedures in the genetic engineering studies and industries involved with PCR can be speeded up.
    • 一种DNA合成方法,其通过聚合酶链式反应(PCR)进行DNA合成所需的时间缩短,其特征在于使用DNA聚合酶,其量有效提供超过10ng每50ul约2kb的扩增DNA片段 的反应混合物,当在以下条件(A)和(B)下进行PCR时:(A)反应混合物:将50体积的包含DNA聚合酶的反应混合物,1ng来自大肠杆菌的基因组DNA和10ng 各引物Eco-1和Eco-2(引物Eco-1和Eco-2的核苷酸序列分别示于序列表的SEQ ID NO:10和11中) 并具有适合于DNA聚合酶的组合物; 和(B)反应条件:35个循环的PCR,其中一个循环由99℃,1秒-66℃,7秒; 用于DNA合成方法的DNA合成试剂盒; 以及PCR试剂的制造。 根据本发明,可以加快基因工程研究和涉及PCR的行业的程序。