扩增产物的毛细管电泳分离 ( Amplification of capillary electrophoresis separation )

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Presentation transcript:

扩增产物的毛细管电泳分离 ( Amplification of capillary electrophoresis separation ) 钟树荣（PhD，副教授）昆明医科大学法医学院

In the second quarter Amplification of capillary electrophoresis separation

Electrophoresis (electrophoresis) is to point to in electrolyte solution, under the action of charged particles in the applied electric field, towards its opposite charge electrode at the speed of different mobile phenomenon. Using electrophoresis separation and analysis of some of the chemical or biological material or purification method and the technology is called electrophoresis.

High performance capillary electrophoresis (HPCE)

The characteristics of high performance capillary electrophoresis apparatus 1. The PCR products need pretreatment before electrophoresis 2. Automatic glue before electrophoresis 3. Electric injection mode 4. High voltage power supply (15000 v) 5. Capillary column 6. The separation medium: viscous polymer solution 7. The fluorescent detection system 8. Automated detection and analysis

The PCR product of pretreatment PCR products before capillary electrophoresis, ten times the size of formamide, make DNA denaturation. Denaturant urea and formamide or DNA bases in the hydrogen bond formation, prevent the formation of complementary double-stranded DNA. Formamide processed DNA single-strand DNA molecules.

Formamide: LIZ500 = per molecular weight (internal standard) = 9:1 formamide LIZ mixture: PCR products 95 ℃ and 0 ℃ (ice)

High performance capillary electrophoresis (HPCE)

ABI 3100 16-capillary array Glue pump system

Electric injection way

Close-up of ABI Prism 310 Sample Loading Area

Capillary column ABI 3100 16-capillary array ABI Capillary column

Capillary viscosity of polymer solution, forming a certain aperture of mesh structure, has a molecular sieves. Denatured DNA fragments in capillary electrophoresis electrophoresis mobility and fragment size into a linear relationship.

The advantages of capillary electrophoresis Apparatus is simple and easy to automate Fast analysis speed, high separation efficiency Sample quantity is little, high sensitivity A wide range of application

Electrophoresis the whole process To prepare programming Start the Pump the glue sample Electrophoretic separation Collect signal Processing the signal The data analysis