Polymerase enzymes and reagents for enhanced nucleic acid...

C - Chemistry – Metallurgy – 12 – N

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C12N 9/12 (2006.01) C12N 15/00 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2666517

Compositions that include DNA polymerases having increased residence times for nucleotide analogues, particularly modified recombinant .PHI.29-type DNA polymerases with such increased residence times, are provided. Methods of making the polymerases and of using the polymerases in sequencing and DNA amplification are also provided. Compositions including .alpha.-thiophosphate nucleotide analogues with four or more phosphate groups are described, as are methods for determining the sequence of nucleic acid molecules using such analogues.

La présente invention concerne des compositions comprenant des ADN polymérases présentant des temps de séjour accrus pour des analogues nucléotidiques , notamment des ADN polymérases de type .PHI.29-type recombinantes modifiées avec de tels temps de séjour accrus. L'invention concerne également des procédés de fabrication des polymérases et d'utilisation des polymérases dans le séquençage et l'amplification d'ADN. L'invention concerne en outre des analogues nucléotidiques .alpha.-thiophosphate avec au moins quatre groupes phosphates, ainsi que des procédés pour déterminer la séquence de molécules d'acide nucléique au moyen de tels analogues.

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