C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) B01L 3/14 (2006.01) B01L 7/00 (2006.01) C07H 21/00 (2006.01) G01N 21/25 (2006.01) G01N 21/64 (2006.01) G01N 21/03 (2006.01) G01N 21/07 (2006.01) G01N 35/00 (2006.01) G01N 35/02 (2006.01)
Patent
CA 2257109
Methods of monitoring hybridization during polymerase chain reaction are disclosed. These methods are achieved with rapid thermal cycling and use of double stranded DNA dyes or specific hybridization probes. A fluorescence resonance energy transfer pair comprises fluorescein and Cy5 or Cy5.5. Methods for quantitating amplified DNA and determining its purity are carried out by analysis of melting and reannealing curves.
L'invention concerne des méthodes pour contrôler l'hybridation pendant la réaction en chaîne de la polymérase. Ces méthodes sont mises en oeuvre au moyen d'un cyclage thermique rapide et de l'utilisation de colorants d'ADN double brin ou de sondes d'hybridation spécifiques. Une paire d'oligonucléotides de transfert énergétique de résonance à fluorescence comprend de la fluorescéine et du Cy5 ou Cy5.5. Des méthodes pour quantifier l'ADN amplifié et déterminer sa pureté sont réalisées par analyse des courbes de fusion et de ré-annelage.
Rasmussen Randy P.
Ririe Kirk M.
Wittwer Carl T.
Smart & Biggar
University Of Utah Research Foundation
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