http://rdf.ncbi.nlm.nih.gov/pubchem/patent/ES-2354745-T3
Outgoing Links
| Predicate | Object |
|---|---|
| assignee | http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_3d77625451c3066a1c9a475578e417bc |
| classificationCPCAdditional | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6818 |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6823 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6827 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12Q1-68 |
| filingDate | 2007-09-24-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| grantDate | 2011-03-17-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| inventor | http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_1d691aa804cd0ac52bfa990386f614d9 http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_501add0443f1dca27b4ed28548c2d337 http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_04d581276e5574585b43ee9dd969b491 |
| publicationDate | 2011-03-17-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | ES-2354745-T3 |
| titleOfInvention | METHOD FOR DETECTION OF MUTANT ALELOS COMBINING REAL-TIME PCR AND REMS-PCR. |
| abstract | Method for detecting the presence or absence of a mutant DNA sequence in a sample, the method comprising the steps of: a) contacting the sample with an oligonucleotide system in hybridization conditions to form a reaction mixture, including said oligonucleotide system a reverse primer and a mutagen primer, wherein i) said reverse primer includes: - a region of the 5 'end comprising a recognition sequence that can be cut by a high temperature resistant restriction endonuclease; said region of the 5 'end having a coupled detection system covalently attached at its ends, so that, when the recognition sequence is cleaved by said high temperature resistant restriction endonuclease, a signal is generated; - a region of the 3 'end capable of hybridizing with a complementary region downstream of the supposed mutant DNA sequence; ii) said mutagenic primer comprises: a region of the 5 'end comprising a first recognition sequence that can be cut by a high temperature resistant restriction endonuclease; - a region of the 3 'end capable of hybridizing with a complementary region of the opposite end of the other chain of the sample DNA sequence, said mutagenic primer has a sequence so that a second recognition sequence is created that can be cut by the high temperature resistant restriction endonuclease, only when said primer is extended in the wild type sequence so that the restriction endonuclease digests the amplicon and suppresses the amplification reaction; and said second recognition sequence that can be cut by high temperature resistant restriction endonuclease does not polymerize when the mutant sequence is present in the DNA sample; b) add, with appropriate substrates and cofactors, in an ionic environment and of suitable pH, both a temperature resistant DNA polymerase and said high temperature resistant restriction endonuclease to said reaction mixture under predetermined reaction conditions, such that, if the mutant DNA sequence is present in the sample, said reverse primer and said mutagen primer hybridize therewith and prime the DNA polymerase reaction to obtain a first specific amplified product; c) cycling the hybridization of said oligonucleotide system so that a second specific amplified product is extended comprising the 5 'end region of the reverse primer that forms a double chain recognition site for said restriction endonuclease so that the endonuclease High temperature resistant restriction cuts specifically in the recognition sequence and induces the generation of a signal by the coupled detection system; d) detect the generated signal. |
| priorityDate | 2006-09-26-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
Total number of triples: 30.