Claims
- 1. A primer having a 3′ portion complementary to a target nucleic acid sequence adjacent to at least one specific nucleotide which may or may not be present in a sample and a 5′ portion complementary to a preselected nucleotide sequence.
- 2. A method for detecting the presence or absence of a target nucleic acid sequence in a sample, said method comprising
(i) adding to said sample an oligonucleotide primer to provide a primer extension product including said target sequence, if present, in said sample, said primer having
a 3, end portion complementary to a portion of said target sequence immediately adjacent at least one specific nucleotide to provide an extension product of said primer which includes said specific nucleotide, and a 5′ end portion complementary to a preselected nucleic acid sequence different from said target sequence; (ii) adding to the product of step (i) a DNA polymerase and one or more labeled deoxynucleoside triphosphates to provide a labeled extension product of said primer which includes said target sequence and said specific nucleotide if present; (iii) subjecting said labeled primer extension product to hybridization conditions with a preselected nucleic acid sequence complementary to the 5′ end portion of said primer; and (iv) determining whether hybridization occurs.
- 3. The method according to claim 2 wherein said sample may contain a plurality of target sequences and wherein said sample is subjected to polymerase chain reaction amplification with at least one pair of oligonucleotide primers for two or more of said target sequences.
- 4. The method according to claim 2 wherein said nucleic acid target sequence$ are alleles of one another, the 3′ portion of said primer is positioned immediately adjacent to the variant nucleotide responsible for the allelism, and said primer extension reactions are independently performed with each of the four deoxynucleoside triphosphates such that only one primer extension reaction will occur for each of said alleles, said deoxynucleoside triphosphate being labeled whereby a single labeled primer extension product is produced-for each of said alleles.
Priority Claims (1)
Number |
Date |
Country |
Kind |
PCT/US92/05133 |
Jun 1992 |
US |
|
Government Interests
[0001] This invention was made with government support under Grant No. HG00099 awarded by the National Institutes of Health. The government has certain rights in the invention.
Continuations (3)
|
Number |
Date |
Country |
Parent |
09661384 |
Sep 2000 |
US |
Child |
09860761 |
May 2001 |
US |
Parent |
09413790 |
Oct 1999 |
US |
Child |
09661384 |
Sep 2000 |
US |
Parent |
08193039 |
Feb 1994 |
US |
Child |
09413790 |
Oct 1999 |
US |