Claims
- 1. A method of detecting, in a sample containing a mixture of double stranded DNA polynucleotides, a target polynucleotide region having a sequence identical to a defined sequence, comprising
- reacting the sample with a peptide nucleic acid (PNA), having a sequence complementary to said defined sequence, under conditions that allow the formation of a complex between perfectly matched PNA and target-region sequences, but not between imperfectly matched PNA and target-region sequences having a single base pair mismatch, where said reacting includes contacting said sample with said PNA at about room temperature, and subsequently elevating the temperature of the reaction to between about 50.degree. C. and about 80.degree. C. to achieve conditions that allow the formation of complexes between perfectly matched PNA and target-region sequences, but not between said imperfectly matched PNA and target-region sequences
- separating complexed from non-complexed polynucleotides in the sample and
- detecting the presence of said complex,
- wherein said PNA is immobilized on a solid support, said reacting includes incubating the sample with the support, said separating includes washing said support to remove unbound polynucleotides, and said detecting includes detecting support-bound polynucleotides and adding labeled polycationic reporter.
- 2. A method of detecting, in a sample containing a mixture of polynucleotides, a target polynucleotide region having a sequence identical to a defined sequence, comprising
- reacting the sample with a peptide nucleic acid (PNA), having a seguence complementary to said defined sequence, under conditions that allow the formation of a complex between perfectly matched PNA and target-region sequences, but not between imperfectly matched PNA and target-region sequences,
- separating complexed from non-complexed polynucleotides in the sample by capillary electrophoresis, wherein said capillary electrophoresis instrument is a free-solution capillary electrophoresis instrument; and
- detecting the presence of said complex.
- 3. The method of claim 2, wherein said separating is carried out at a temperature between about 50.degree. C. and 95.degree. C.
- 4. The method of claim 3, wherein said separating is carried out at a temperature between about 50.degree. C. and about 80.degree. C.
- 5. The method of claim 2, wherein said detecting includes detecting using a UV-absorbance detector.
- 6. The method according to claim 2, wherein said PNA is at least 10 nucleotides in length.
- 7. The method of claim 6, wherein said PNA is not more than 15 nucleotides in length.
Parent Case Info
This application claims priority under 35 U.S.C. 120 to provisional patent application Ser. No. 60/013,913, filed Mar. 21, 1996, incorporated herein by reference.
Government Interests
This work was supported in part by grant number MH 45423-06 awarded by the National Institute of Mental Health. Accordingly, the United States Government has certain rights in this invention.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
5593559 |
Wiktorowicz |
Jan 1997 |
|
Foreign Referenced Citations (1)
Number |
Date |
Country |
WO 9220703 |
Nov 1992 |
WOX |