Method for colourimetric identifying detecting DNA molecule

A DNA molecule and colorimetric technology, applied in the field of detection of DNA molecules, has broad application prospects, simple operation, and the effect of quantitative detection

Inactive Publication Date: 2005-02-23
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on the development of color-changing molecular ordered assemblies with DNA molecular recognition and detection functions by using polymerized diacetylene lipid films

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Probe DNA1: 5'-TCTCAACTCGTATTTTTT-cholesteryl-3'

[0032] Probe DNA2: 5'-cholesteryl-TTTTTTCGCATTCAGGAT-3'

[0033] cDNA: 5'-TACGAGTTGAGAATCCTGAATGCG-3'

[0034] mDNA: 5'-GCGTAACTCCTAAGAGTTGAGCTA-3'

[0035] mDNA1: 5'-TACCTCTACAGAATCCTGAATGCG-3'

[0036] mDNA2: 5'-TAGCTGTTGAGAATCCTGAATGCG-3'

[0037] mDNA3: 5'-TAGGAGTTGAGAATCCTGAATGCG-3'

[0038] Preparation of vesicle 1: (1), probe DNA1 (mole percent 10%), 10,12-tricosyldiynoic acid or 10,12-pentacosyldiynoic acid (mole percent 70%) %) and a chloroform solution of phospholipids (20% by mole%) are mixed, and the organic solvent is evaporated by rotary evaporation; (2), adding distilled water or buffer solution (pH=7.4), so that the molar concentration of diacetylene lipid is 1mM; ( 3), ultrasonically disperse at 60°C for 55min; (4), store the dispersion obtained in (3) overnight at 4°C; Irradiate and polymerize for 10 minutes to obtain blue polymerized diacetylene vesicles 1 functionalized with probe DNA molecules...

Embodiment 2

[0045] Probe DNA 1: 5’-TCTCAACTCCGGTTTTTT-cholesteryl-3’

[0046] Probe DNA 2: 5’-cholesteryl-TTTTTTAAGATTCAGGAT-3’

[0047] cDNA: 5'-CCGGAGTTGAGAATCCTGAATTTC-3'

[0048] mDNA: 5'-GCGTAACTCCTAAGAGTTGAGCTA-3'

[0049] mDNA1: 5'-CACCAGTAGAGAATCCTGAATTTC-3'

[0050] mDNA2: 5'-CCTTAGTTGAGAATCCTGAATTTC-3'

[0051] mDNA3: 5'-CTGGAGTTGAGAATCCTGAATTTC-3'

[0052] Preparation of vesicle 1: (1), probe DNA 1 (5% by mole), 5,7-tricosyldiynoic acid or 5,7-pentacosyldiynoic acid (5% by mole 50%) and a chloroform solution of phospholipids (45% by mole percent) were mixed, and the organic solvent was evaporated by rotary evaporation; (2), distilled water or buffer solution (pH=9) was added so that the molar concentration of diacetylene lipid was 1 mM; (3), ultrasonically disperse at 50°C for 30min; (4), store the dispersion obtained by (3) overnight at 4°C; Light irradiated and polymerized for 10 minutes to obtain blue polymerized diacetylene vesicles 1 functionalized with probe DNA mol...

Embodiment 3

[0059] Probe DNA 1: 5’-GGTTAACTCGTATTTTTT-cholesteryl-3’

[0060] Probe DNA 2: 5’-cholesteryl-TTTTTTCGCATTCAGGCC-3’

[0061] cDNA: 5'-TACGAGTTAACCGGCCTGAATGCG-3'

[0062] mDNA: 5'-GCGTAACTCCTAAGAGTTGAGCTA-3'

[0063] mDNA1: 5'-TAGATGTCCACCGGCCTGAATGCG-3'

[0064] mDNA2: 5'-TACGAGTTAACCGGCCTGAATGCG-3'

[0065] mDNA3: 5'-TACCAGTTAACCGGCCTGAATGCG-3'

[0066] Preparation of vesicle 1: (1), probe DNA1 (2% by mole), 10,12-tricosyldiynoic acid or 10,12-pentacosyldiynoic acid (98% by mole %) chloroform solutions were mixed, and the organic solvent was evaporated by rotary evaporation; (2), distilled water or buffer solution (pH=5) was added, so that the molar concentration of the diacetylene lipid was 1 mM; (3), under the condition of 40° C. Ultrasonic dispersion for 20 minutes; (4), the dispersion obtained by (3) was stored overnight at 4°C; (5) the dispersion of (4) was irradiated with 256nm ultraviolet light for 10 minutes to obtain probe DNA molecules 1 functionalized blue p...

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Abstract

A colorimetric identifying and detecting DNA method is disclosed. It has double functions of allochroic indicant group and reinforced allochroism by functional polymeric butadiyne vesiculation. It achieves simple and rapid process.

Description

technical field [0001] The invention belongs to biomolecule detection methods, in particular to a method for detecting DNA molecules. Background technique [0002] Commonly used methods for identifying and detecting DNA molecules include radioactive labeling, fluorescent labeling, enzyme labeling, and chemiluminescent methods. Most of these methods involve some expensive detection instruments and equipment, and there is also the possibility of radioactive contamination for radioactive labels. Therefore, the development of radiation-free background, simple operation, rapid, economical, simple equipment or no instrument [0003] New methods for the identification and detection of DNA molecules in organotypic devices have important practical significance. [0004] In recent years, the research on molecular ordered assemblies has developed rapidly, which provides a favorable tool for the use of bionic technology to study new methods with biomolecular recognition. Among them, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 马占芳王春刚
Owner NORTHEAST NORMAL UNIVERSITY
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