CN1526828A - MGB marked DNA molecule beacon probe and its application - Google Patents
MGB marked DNA molecule beacon probe and its application Download PDFInfo
- Publication number
- CN1526828A CN1526828A CNA031071813A CN03107181A CN1526828A CN 1526828 A CN1526828 A CN 1526828A CN A031071813 A CNA031071813 A CN A031071813A CN 03107181 A CN03107181 A CN 03107181A CN 1526828 A CN1526828 A CN 1526828A
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- China
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- probe
- mgb
- target dna
- dissolving temperature
- point mutation
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Abstract
DNA double strand small groove binding group DP13 is used as molecular mark of oligonucleotide and fluorescent probe 3 end is used as molecule beacon in the detection of gene point mutation or mononucleotide polymorphism. The small groove binding group can bind with the ends of probe and target DNA hybrid strand to raise the double strand dissolving temperature and stability of the hybrid strand. In the case of unchanged dissolving temperature, the number of probes and target DNA complementing bases may be reduced properly to reduce probe synthesizing cost, increase the difference between normal probe and mutant probe in dissolving temperature and raise the sensitivity and accuracy in detecting gene point mutation and SNP obviously.
Description
Technical field: biomedical gene molecule diagnostic techniques
Technical background:
Molecular beacon (molecular beacon) is the translations molecular beacon again, it is a kind of special fluorescent mark oligonucleotide probe, can be used for detecting in real time polymerase chain reaction (PCR) product quantity and efficient, have highly sensitive, specificity good, advantage that can qualitative, quantitative.The principle that molecular beacon is used for gene diagnosis is: at 5 ' and 3 ' end difference mark report fluorophor (reporter of oligonucleotide probe, R) and quenching group (quencher, Q), and make several bases that R and two groups of Q close on (4-6) complementation, intermediary sequence then be can with target DNA specificity complementary base sequence.Under free situation, the two ends complementation of fluorescent probe is folded into loop-stem structure (stem-loop structure) like this.Very approaching on R group and the Q group space, the fluorescent energy that the R group penetrates after exciting can be absorbed cancellation by the Q group, does not produce fluorescent signal.And after probe and target DNA formed the hybridization chain, the more intermediate annular part base of probe combined with the target DNA complementation, forced self complementary two strands of probe stem to be opened, and R group Q group space length widens, and can launch fluorescent signal after being excited.Adopt common Q group mark, still can form the hybridization chain with target DNA in order to guarantee fluorescent probe in the pcr amplification process, probe must surpass PCR upstream and downstream primer length, usually greater than 18 bases.If wherein after the change of one, two base, the difference of the melting temperature (Tm) of its heteroduplex (Tm) is less above 18 bases but probe and target DNA are complementary, the sensitivity that detects single base mutation or polymorphism (SNP) is not high.
Summary of the invention:
Present technique adopts dna double chain ditch, and (minorgroovebinder, MGB) DPI3 (dihydrocyclopyrroloindole tripeptide) is marked on 3 ' end of fluorescent probe as quenching group (Q) in conjunction with gene.MGB can combine in the two strands of 5-6 base pair of probe and target DNA hybridization chain end, has improved the Tm value and the stability of hybridization chain.Keeping under the constant situation of Tm value, can suitably reduce in the middle of the fluorescent probe and target DNA complementary base number, can foreshorten to 13-16 base usually.So not only can reduce the synthetic cost of fluorescent probe, and increase the difference of Tm value between normal probe and the mutant probe, improve sensitivity and accuracy rate that point mutation and SNP detect.
Claims (3)
1,3 ' of oligonucleotide probe end mark ditch conjugated group (MGB) DPI3, fluorescent substances such as 5 ' end glimmering group FAM.TET of mark or Cy3.
2, the specificity complementary sequence through MGB and fluorescently-labeled oligonucleotide probe and target gene contains 13-16 base length.
3, MGB labeled oligonucleotide fluorescent probe is applied to the qualitative and quantitative analysis of point mutation, single nucleotide polymorphism (SNP) and gene.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031071813A CN1526828A (en) | 2003-03-07 | 2003-03-07 | MGB marked DNA molecule beacon probe and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031071813A CN1526828A (en) | 2003-03-07 | 2003-03-07 | MGB marked DNA molecule beacon probe and its application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1526828A true CN1526828A (en) | 2004-09-08 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA031071813A Pending CN1526828A (en) | 2003-03-07 | 2003-03-07 | MGB marked DNA molecule beacon probe and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1526828A (en) |
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2003
- 2003-03-07 CN CNA031071813A patent/CN1526828A/en active Pending
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |