Method for quantitative detection of DNA using quantum dot as fluorescence tracer

A fluorescent tracer, quantitative detection technology, applied in the field of quantitative detection of DNA, to achieve the effect of a simple DNA quantitative detection method

Inactive Publication Date: 2006-12-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

We believe that quantum dots can also be used in the quantitative detection of DNA. So far, there has been no relevant literature report, and our related research articles have been submitted.

Method used

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  • Method for quantitative detection of DNA using quantum dot as fluorescence tracer
  • Method for quantitative detection of DNA using quantum dot as fluorescence tracer

Examples

Experimental program
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Embodiment 1

[0012] Dissolve cadmium chloride in an alkaline (pH10) aqueous solution containing thioglycolic acid, add ionic tellurium source NaHTe (prepared from tellurium powder and sodium borohydride), and react at 100°C for 30 to 240 minutes. The reaction time is different to obtain quantum dots. The particle size is different, the color of the solution is from green to orange and finally red, and CdTe quantum dots with carboxyl groups on the surface are synthesized. CdTe quantum dots with different particle sizes in the range of 1.6nm to 3nm can be obtained by this method. Here, quantum dots with a particle size of 3 nanometers are selected and excited at any wavelength less than 500 nanometers to obtain fluorescence with an emission wavelength of 560 nm.

[0013] The full-length genome is the 93.857kb breast cancer-associated antigen gene (brcaa1 gene), its gene bank acceptance number is AF208045, and a series of different concentrations (1.25, 2.5, 5, 10, 20, 40ng / mL) of brcaa1 are p...

Embodiment 2

[0015] Using cadmium chloride, tellurium powder, and sodium borohydride as raw materials, in an acidic (pH4) aqueous solution containing cysteine, reflux reaction for 30 to 240 minutes to synthesize CdTe quantum dots with amino and carboxyl groups on the surface, by controlling the reaction Time can prepare CdTe quantum dots with different particle sizes (2.8nm-4.5nm), and the obtained CdTe quantum dots' ultraviolet absorption peak position redshifts from 531nm to 590nm, and the fluorescence emission peak redshifts from 552nm to 600nm. Here, quantum dots with a particle size of about 4 nanometers are selected, excited at any wavelength less than 500 nanometers, and fluorescence with an emission wavelength of 582 nm can be obtained.

[0016] The full-length genome is the 93.857kb breast cancer-associated antigen gene (brcaa1 gene), its gene bank acceptance number is AF208045, and a series of different concentrations (5, 10, 20, 40, 80, 160 μg / mL) of brcaa1 are prepared with dist...

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Abstract

A quantitative determination method for DNA with quantum point as fluorescent tracer belongs to the DNA detecting domain. The invention builds the standard detection curve making use of that the quantum point has excellent fluorescent characteristic, DNA can quench the fluorescent signal of the quantum point and the signal quenched degree and the DNA quantity construct positive correlation, and obtains the DNA concentration of the solution using the fluorescent intensity and the standard curve of the quantum point in the measuring solution; the signal quenched degree and the DNA quantity construct positive correlation and the specific numerical relationship is: DNA concentration equals to 10(1264.4-Y)/1786ng/mL; the DNA concentration of the solution can be calculated using the fluorescent intensity and the standard curve of the quantum point in the measuring solution. The invention can be used in calculation of fluorescence labeling DNA quantity, and the method is simple, fast and sensitive.

Description

technical field [0001] The invention relates to a method in the technical field of DNA detection, in particular to a method for quantitatively detecting DNA using quantum dots as fluorescent tracers. Background technique [0002] The traditional method for quantitative detection of DNA is spectrophotometry, which detects the absorbance value at 260 nanometers, and the DNA content = OD 260 ×50μg / ml, this method is more complicated to operate; another common method is to use ethidium bromide combined with DNA molecules to generate red fluorescence under the excitation of ultraviolet light. The fluorescence intensity is proportional to the amount of DNA. Using the standard curve, DNA Quantitative analysis, because ethidium bromide is a carcinogen, should not be promoted. Currently, TaqMan probes are used for fluorescent quantitative PCR detection, but the cost is high and the background signal is strong, which has a certain impact on a wide range of quantitative detection appl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 崔大祥高峰贺蓉潘碧峰
Owner SHANGHAI JIAO TONG UNIV
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