Primers and methods for the detection and discrimination of nucleic acids
AssigneeLife Technologies Corporation
InventorsIRINA NAZARENKO, AYOUB RASHTCHIAN
Life of this patent
Estimating the term…
Abstract
The present invention provides novel primers and methods for the detection of specific nucleic acid sequences. The primers and methods of the invention are useful in a wide variety of molecular biology applications and are particularly useful in allele specific PCR.
Claims22
A composition for quantifying or detecting one or more target nucleic acid molecules in a sample comprising one or more detectably labeled oligonucleotides and one or more target nucleic acid molecules to be detected or quantified, wherein said oligonucleotides comprise one of more detectable labels located internally and/or at or near the 3′ and/or 5′ termini of said oligonucleotides and wherein said label undergoes a detectable change in an observable property upon becoming part of a double stranded molecule.
11-17. (canceled)
A method for amplifying a double stranded nucleic acid molecule, comprising:
- providing a first and second primer, wherein said first primer is complementary to a sequence within or at or near the 3′-termini of the first strand of said nucleic molecule and said second primer is complementary to a sequence within or at or near the 3′-termini of the second strand of said nucleic acid molecule;
- hybridizing said first primer to said first strand and said second primer to said second strand in the presence of one or more of the polymerases, under conditions such that a third nucleic acid molecule complementary to all or a portion of said first strand and a fourth nucleic acid molecule complementary to all or a portion said second strand are synthesized;
- denaturing said first and third strand, and said second and fourth strands;
- and
- repeating the above steps one or more times, wherein one or more of the primers comprise a detectable label internally and/or at or near its 3′ and/or 5′ termini and/or comprises one or more hairpin structures.
20-47. (canceled)
A method of determining the presence of at least one particular nucleotide of interest at a specific position in a target nucleic acid molecule, comprising:
- providing at least one target nucleic acid molecule having said nucleotide of interest at a specific position;
- contacting said target nucleic acid molecule with at least one oligonucleotide, wherein at least a portion of the oligonucleotide is capable of forming base pairs or hybridizing with at least a portion of the nucleic acid molecule and wherein the oligonucleotide comprises at least one specificity enhancing group and/or at least one label;
- and
- contacting the oligonucleotide and the target nucleic acid molecule with a polymerase less able to extend the oligonucleotide when the 3′-most nucleotide of the oligonucleotide does not base pair with the target nucleic acid and more able to extend the oligonucleotide when the 3′-most nucleotide of the oligonucleotide base pairs with the target nucleic acid molecule.
(canceled)
Dependencies are inferred from the claim text.