Capillary array electrophoresis detection method based on spatial correction and spectral correction

Abstract

The invention provides a capillary array electrophoresis detection method based on spatial correction and spectral correction, and the method comprises the following steps: (1) utilizing a Raman spectrum to position imaging positions of all capillaries in a capillary array on a charge coupled element; (2) performing spectrum unscrambling on multi-color fluorescent dyes in each capillary through the spatial correction; and (3) performing analysis on DNA (deoxyribonucleic acid) fragments of electrophoresis of the capillary array. In the method, the Raman spectrum of laser excitation capillary quartz or solvent water in the capillaries is adopted for realizing spectral imaging, thereby realizing the position positioning and parallelism adjustment of the capillaries on a CCD (charge coupled device); a fluorescent dye combined matrix is adopted for realizing simultaneous detection of the multi-color fluorescent dyes in the single capillary and the pixel combination way is adopted for improving the stability of a distribution matrix, reducing the amount of calculation and improving the detection stability; and a device for implementing the method is simple in structure and easy to operate.

Description

technical field [0001] The invention relates to a capillary array electrophoresis detection method, which belongs to the technical field of high-throughput microanalysis. Background technique [0002] DNA sequencing is an important technology in modern molecular biology research, and electrophoresis technology is an indispensable technical means in DNA sequencing. Because the capillary has good heat dissipation performance, it is allowed to apply a voltage as high as 30kV at both ends of the capillary, and the longitudinal electric field strength of the separation capillary can reach more than 400V / cm, so the separation operation can be completed in a short time, reaching a very high Separation efficiency (the number of theoretical plates reaches more than 400000 / m, up to 107 / m order of magnitude). Because the inner diameter of the capillary is very small (generally <100 μm), for a capillary with an inner diameter of 50 μm and a length of 50 cm, the volume is less than 1...

AI Technical Summary

Problems solved by technology

However, due to the uncertainty of the positional relationship between the capillaries, there are errors in the qualitative and quantitative analysis of micro reagents by the capillary array.

In addition, for the simultaneous detection of multicolor fluorescent dyes in a single capillary, since the fluorescent bands of the fluorescent markers are relatively wide, resulting in a large overlap between different fluorescent spectra, it is difficult to distinguish them well through ordinary spectroscopy. Also caused a large error in capillary electrophoresis analysis

At present, there is no mature technology to eliminate these errors

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