DEAE-Dextran method.pptx

DEAE-Dextran method

1. 1

2. DEAE-Dextran method

3. Content DEAE-Dextran Principle DEAE-dextran method Example “ mammalian cultured cells” Advantages and disadvantages Conclusion 3

4. 4

5. DEAE-Dextran Diethylaminoethyl-dextran It is a polycationic derivative of the carbohydrate polymer dextran. I t is one of the first chemical reagents used to transfer nucleic acids into cultured mammalian cells. (Vaheri and Pagano, 1965). 5

6. 6

7. steps of deae-dextran Step:1 Mix nucleic acid with DEAE Step:2 Add Nucleic Acid DEAE Step:3 Induce The Uptake 7 Step :4 Wash Cells Step:5 Assay Cells

8. STEP NO: 1 Mix Nucleic Acid With DEAE Mix nucleic acid with DEAE-dextran solution in transfection medium or phosphate-buffered saline solution. Nucleic acid-DEAE-dextran complexes are formed via electrostatic interactions between the polymer and phosphate backbone of the nucleic acid. 8

9. STEP NO: 2 Add the nucleic acid deae Add the nucleic acid-DEAE-dextran complexes to the cells, which adhere to the cell surface via electrostactic interactions. 9

10. STEP NO: 3 Induce The Uptake Induce the uptake of the nucleic acid-DEAE-dextran complexes by osmotic shock using DMSO or glycerol. 10

11. STEP NO: 4 Wash cells Wash cells to remove the complexes and incubate to allow gene expression. 11

12. STEP NO: 5 Assay cells Assay cells for transient gene expression. 12

13. 13

14. 14

15. Advantages Disadvantages Low Cost Easy Technique Quick process Can be used for wide variety of cells High conc of DEAE dextran can be toxic Efficiency of transfection is low Uptake of genes is also less compared to other techniques 15

16. CONCLUSION DIETHYLAMINOETHYL – dextran Cationic Nature (+) DEAE + DNA +ive Electrostatic interaction / Cytoplasm Cell transformed 16

17. 17