United States Patent
US Patent 6033854: Quantitative PCR using blocking oligonucleotides
US 6033854 · granted 2000-03-07
Abstract
Disclosed is a method for determining the number of copies of a target nucleic acid relative to the number of copies of a reference nucleic acid in a biological specimen. The method includes amplifying the target nucleic acid, measuring the amplification of the target nucleic acid, amplifying the reference nucleic acid, measuring the amplification of the target nucleic acid, and comparing the amplification of the target nucleic acid sequence to the amplification of the reference nucleic acid sequence.
| Patent Number | 6033854 |
|---|---|
| Title | Quantitative PCR using blocking oligonucleotides |
| Filed | 1998-01-27 |
| Granted | 2000-03-07 |
| Inventor(s) | Wen, Kurnit; David M., Ning J. |
| Assignee | Biotronics Corporation |
| CPC Classification | C12Q 1/68 |
| Number of Claims | 25 |
Abstract
Disclosed is a method for determining the number of copies of a target nucleic acid relative to the number of copies of a reference nucleic acid in a biological specimen. The method includes amplifying the target nucleic acid, measuring the amplification of the target nucleic acid, amplifying the reference nucleic acid, measuring the amplification of the target nucleic acid, and comparing the amplification of the target nucleic acid sequence to the amplification of the reference nucleic acid sequence.
Claim 1
A method for determining the number of copies of a target nucleic acid relative to the number of copies of a reference nucleic acid in a biological specimen, which method comprisesthe steps of: (a) amplifying the target nucleic acid in said specimen with a polymerase, a first primer specific for said target nucleic acid with or without a segment noncontiguous to a first priming sequence, a second primer specific for said target nucleicacid with or without a segment noncontiguous to a second priming sequence in the presence of a first oligonucleotide which is incapable of acting as a primer for said polymerase, wherein said first oligonucleotide has at least 5 consecutive nucleotidesfully complementary to at least 5 consecutive nucleotides of said first primer; (b) measuring the amplification of the target nucleic acid; (c) amplifying the reference nucleic acid in said specimen with a polymerase, a third primer specific for said reference nucleic acid with or without a segment noncontiguous to a third priming sequence, and a fourth primer specific for saidreference nucleic acid sequence with or without a segment noncontiguous to a fourth priming sequence in the presence of a second
Claims
25 totalA method for determining the number of copies of a target nucleic acid relative to the number of copies of a reference nucleic acid in a biological specimen, which method comprisesthe steps of: (a) amplifying the target nucleic acid in said specimen with a polymerase, a first primer specific for said target nucleic acid with or without a segment noncontiguous to a first priming sequence, a second primer specific for said target nucleicacid with or without a segment noncontiguous to a second priming sequence in the presence of a first oligonucleotide which is incapable of acting as a primer for said polymerase, wherein said first oligonucleotide has at least 5 consecutive nucleotidesfully complementary to at least 5 consecutive nucleotides of said first primer; (b) measuring the amplification of the target nucleic acid; (c) amplifying the reference nucleic acid in said specimen with a polymerase, a third primer specific for said reference nucleic acid with or without a segment noncontiguous to a third priming sequence, and a fourth primer specific for saidreference nucleic acid sequence with or without a segment noncontiguous to a fourth priming sequence in the presence of a second