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  • 标题:PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates
  • 本地全文:下载
  • 作者:W M Barnes
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:1994
  • 卷号:91
  • 期号:6
  • 页码:2216-2220
  • DOI:10.1073/pnas.91.6.2216
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:A target length limitation to PCR amplification of DNA has been identified and addressed. Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased. These improvements were achieved by the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified to high yields from 1 ng of lambda DNA template.
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