Polymerase extension at 3' terminus of pna-dna chimera

Details Polymerase extension at 3' terminus of pna-dna chimera Polymerase extension at 3' terminus of pna-dna chimera

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2381098

The invention provides methods and a kit for primer extension of PNA-DNA chimera from template nucleic acids using polymerases, nucleotide 5'- triphosphates, and primer extension reagents. Structural requirements of the chimera for primer extension include 5 to 15 contiguous PNA monomer units, 3 or more contiguous nucleotides, and a 3' hydroxyl terminus. The chimera and/or a nucleotide is labelled with the fluorescent dyes or other labels. The methods include DNA sequencing, DNA fragment analysis, reverse transcription, mini-sequencing, chromosome labelling, amplification, and single nucleotide polymorphism (SNP) detection.

La présente invention concerne des procédés et une trousse pour l'extension d'amorces de chimère de PNA-ADN, à partir d'acides nucléiques matrice, par utilisation de polymérases, de triphosphates 5' nucléotidiques et de réactifs d'extension d'amorces. Les besoins structuraux de la chimère pour l'extension d'amorces comprennent 5 à 15 unités monomères de PNA contiguës, 3 ou plus nucléotides contigus et une extrémité hydroxyle 3'. La chimère et/ou un nucléotide est marqué avec des colorants fluorescents ou d'autres marqueurs. Les procédés selon l'invention comprennent le séquençage de l'ADN, l'analyse de fragments d'ADN, la transcription inverse, le mini-séquençage, le marquage chromosomique, l'amplification, ainsi que la détection de polymorphismes de nucléotide simple (SNP).

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