Methods and reagents for the polymerase chain reaction amplification of long DNA sequences
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United States of America Patent
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Apr 30, 1996
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app pub date -
Feb 25, 1994
filing date -
Feb 25, 1994
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Abstract
Methods and reagents are provided for the amplification of DNA sequences longer than 10 kilobases by the polymerase chain reaction (PCR). The methods use compositions consisting of a primary thermostable DNA polymerase from Thermus thermophilus combined with a lesser amount of a secondary thermostable DNA polymerase possessing a 3'-to-5' exonuclease activity from Thermococcus litoralis, Pyrococcus species GB-D or Thermotoga maritima. The DNA polymerase compositions, when used with the disclosed reaction buffer, enable amplifications of DNA sequences up to at least 42.2 kilobases in length.
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