Method for rapidly detecting mismatching quantity of bases through capillary electrophoresis

A technology of capillary electrophoresis and capillary, applied in the field of biological analysis, to achieve the effect of high repeatability, simple operation, and expanded application

Active Publication Date: 2015-12-23
CHANGZHOU UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention overcomes the deficiency of detecting the number of mismatched bases in complementary DNA in the prior art, provides a method

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
  • Method for rapidly detecting mismatching quantity of bases through capillary electrophoresis
  • Method for rapidly detecting mismatching quantity of bases through capillary electrophoresis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 1. The capillary adopts a quartz capillary with an inner diameter of 75 μm, a total length of 60 cm, and an effective length of 35 cm. The sample to be tested and the buffer solution flow in the capillary through an 18kV high-voltage power supply.

[0020] 2. (1) Design DNA1-FAM fluorescent probe, which includes two parts of fluorescent dye 6-FAM and DNA1. DNA1 is 5'-GGTTGGTGTGGTTGG-3', and the 3' end is coupled with a fluorescent dye; (2) Design five complementary DNAs, which are 5'-CCAACCACACCAACC-3' (zero mismatched bases), 5'-CCAACCACACCAAC G -3' (one mismatch base), 5'-CCAACCACACCAA GG -3' (two mismatched bases), 5'-CCAACCACACC GGGG -3' (four mismatched bases), 5'-CCAACCACA GGGGGG -3' (six mismatched bases); (3) DNA1-FAM and complementary DNA containing different numbers (0, 1, 2, 4, 6) of mismatched bases at the same concentration ratio 2:2, with an interval of 20s in the capillary, inject samples for 20s respectively, and detect by fluorescence capillary elect

Embodiment 2

[0023] 1-2 steps are the same as in Example 1.

[0024] 3. DNA3 to be detected, the sequence is 5'-CCAA GG ACACCAACC-3', carry out capillary electrophoresis, get S 1 / S Total is 0.2, compared with the standard curve, the number of mismatches detected is 2.

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

PropertyMeasurementUnit
The inside diameter ofaaaaaaaaaa
Full lengthaaaaaaaaaa
Login to view more

Abstract

The invention relates to the technical field of biological analysis, and discloses a method for rapidly detecting the mismatching quantity of bases through capillary electrophoresis. The method comprises the following steps: 1, designing DNA of a 3' end coupling fluorescence dye to form a fluorescence probe; 2, designing several complementary DNAs having mismatching bases with the quantity different from that of the DNA fragment in step 1; and 3, respectively hybridizing the several complementary DNAs with the fluorescence probe in a capillary tube, carrying out fluorescence detection, calculating the peak area ratio (S1/STotal) to obtain a standard curve of the peak area ratio and the quantity of the mismatching bases, hybridizing a sample to be detected with the fluorescence probe in the capillary tube, calculating the electrophoresis peak area, and comparing with the standard curve to determine the quantity of the mismatching bases in the sample to be detected. The method for rapidly detecting the quantity of mismatching bases in the capillary tube has the advantages of simple operation, high repeatability, convenient and fast detection of the quantity of the mismatching bases in the sample, and further development of the application of the DNA fluorescence probe in the field of biological analysis.

Description

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

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
Owner CHANGZHOU 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