Abi prism 3 10 gene analyzer (that is, dna sequencer) uses capillary electrophoresis technology instead of traditional polyacrylamide plate electrophoresis, and uses the company's patented four-color fluorescent dye labeled ddntp (labeled terminator method), so the pcr product generated by single primer pcr sequencing reaction is a single-stranded dna mixture with four different fluorescent dyes at the 3'' end. The difference is 1 base, so that the sequencing pcr products of four fluorescent dyes can be electrophoresed in capillary, avoiding the influence of the mobility difference between lanes and because of the different molecular sizes, the mobility in capillary electrophoresis is different. When molecules pass through the capillary reading window, the ccd (Charge Coupled Device) camera detector in the laser detector window can detect fluorescent molecules one by one. The excited fluorescence is divided by grating to distinguish different colors of fluorescence representing different substrate information, and the images are synchronized on ccd camera. Analysis software can automatically convert different fluorescence into dna sequence, so as to achieve the purpose of dna sequencing. The analysis results can be output in various forms, such as gel electrophoresis map, fluorescence absorption peak map or base arrangement order.
It is an advanced precision instrument for measuring the base sequence, size and quantification of dna fragments by computer automatic control, such as automatic glue filling, automatic sampling and automatic data collection and analysis. Pe also provides gel polymers, including dna sequencing gel (pop 6) and genescan gel (pop 4). The pore size of these gel particles is uniform, which avoids the influence of inconsistent gel preparation conditions on sequencing accuracy. It is mainly composed of capillary electrophoresis device, macintosh computer, color printer and electrophoresis accessories. Computers include software for data collection, analysis and instrument operation. It uses the latest ccd camera detector to shorten dna sequencing to 2.5h hours, and pcr fragment size analysis and quantitative analysis are 10 ~ 40 minutes.
Because the instrument has the functions of dna sequencing, pcr fragment size analysis and quantitative analysis, it can perform dna sequencing, heterozygote analysis, single strand conformation polymorphism analysis (sscp), microsatellite sequence analysis, long fragment pcr, rt-pcr (quantitative pcr) and so on. In addition to routine dna sequencing, it can also carry out single nucleotide polymorphism (snp) analysis, gene mutation detection, hla matching and forensic medicine.