Amplification of long nucleic acid sequences by pcr

Details Amplification of long nucleic acid sequences by pcr Amplification of long nucleic acid sequences by pcr

C - Chemistry, Metallurgy

12

N

C12N 9/12 (2006.01) C12N 15/10 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2143229

Methods and reagents are provided for the amplification of nucleic acid sequences, e.g. 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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