Asymmetric cyanine dye compound and its application

A technology of cyanine dyes and compounds, applied in the field of application of ROX channel fluorescence quenchers

Inactive Publication Date: 2015-12-30
ZHEJIANG UNIV
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the prior art, ROX fluorescent dyes are generally used to adjust the difference between PCR tubes and tubes caused by PCR loading errors. At present, there is no ROX channel quencher that is widely used.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Asymmetric cyanine dye compound and its application
  • Asymmetric cyanine dye compound and its application
  • Asymmetric cyanine dye compound and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: the synthesis of compound (III)

[0034]

[0035] 2,3-Dimethylbenzoxazole p-toluenesulfonate (180mg, 0.52mmol), benzamidine (160mg, 0.8mmol) and 2ml of acetic anhydride were refluxed for 30 minutes, cooled and dropped into ether to obtain Product 170mg.

[0036] Product NMR data 1 HNMR: 1.82 (s, 3H), 2.34 (s, 3H), 6.06 (m, 1H), 7.7 (m, 1H), 4.39 (s, 3H), 7.5-8.6 (m, 13H).

Embodiment 2

[0037] Embodiment 2: the synthesis of compound (II)

[0038]

[0039] 4-Methylquinoline (4.8ml, 0.0363mol) and methyl 2-(4-(bromomethyl)-2-nitrophenyl)propionate (prepared according to NaokiYamakawa et al., Bioorganic & Medicinal Chemistry 19(2011) 3299-3311) (16.44g, 0.05445mol) mixed, react overnight at 110°C, after cooling, add 20ml of methanol to dissolve, then hang and steam to obtain a viscous solid, then add 200ml of acetone and stir, after stirring for 2h, the product precipitates, filter, and dry to obtain the product Compound (II) 6.4g.

[0040] Product NMR data 1 HNMR: 1.51 (s, 3H), 2.66 (s, 3H), 3.6 (d, 3H), 3.7 (m, 1H), 5.95 (s, 2H), 7.5-8.6 (m, 9H).

Embodiment 3

[0041] Embodiment 3: the synthesis of compound (I-1)

[0042]

[0043] Compound III (58mg, 0.13mmol) and compound (II) (89mg, 0.2mmol) were dissolved in 2ml of pyridine, and heated to reflux for 30 minutes. After the reaction was completed, the pyridine was removed by rotary evaporation, and the residue was poured into water, and 5% Hydrochloric acid, filter after the product is precipitated, dissolve the product obtained by filtration in 30ml of methanol and water (volume ratio 1:1), add 80mg of sodium hydroxide for hydrolysis, after hydrolysis, drop concentrated hydrochloric acid again, after the product precipitates, Filter and dry to get the final product.

[0044] Product NMR data 1HNMR: (1.56(m, 3H), 3.82(m, 1H), 4.39(s, 3H), 5.5(m, 3H), 6.65(m, 1H), 6.71(m, 1H), 8.54(m, 1H ), 7.3-8.6 (m, 12H).

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides an asymmetric cyanine dye compound (I) and application of the asymmetric cyanine dye compound serving as a quenching agent. The asymmetric cyanine dye fluorescence quenching agent compound can effectively quench fluorescence of an ROX channel, meanwhile, double DNA strands can be stabilized, the melting temperature in the PCR process is increased by 2 DEG C to 3 DEG C, the compound not only can be applied to a conventional real-time fluorescence quantitative PCR detection method but also can be effectively used in the field of analysis for single nucleotide polymorphism. Please see the formula of the asymmetric cyanine dye compound in the specification.

Description

(1) Technical field [0001] The invention relates to an asymmetric cyanine dye compound and its application as a fluorescence quencher, especially a ROX channel fluorescence quencher. (2) Background technology [0002] Nucleic acid hybridization technology is one of the basic techniques of molecular biology, which can be used for diagnosis of genetic diseases, human body identification, microbial identification, paternity identification, and is gradually being used for specific, sensitive and rapid diagnosis of viruses in recent years. [0003] An important aspect of nucleic acid hybridization techniques is the detection of hybridization. A frequently used tool is the fluorescent oligonucleotide probe. A particularly useful type of fluorescent probe is the self-quenching probe, which includes a reporter The dye and the quencher dye interact through the process of fluorescence resonance energy transfer (FRET). Although the design of different probes using this motif may diffe...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C09B23/06C12Q1/68
Inventor 方维佳郑怡
Owner ZHEJIANG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap