Blotting techniques ppt- finalpptxfinal.pptx

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1. University of Agriculture Faisalabad Assignment of: Clinical Epidemiology Topic: “ Sothren , Westren , and Nothren blotting techniques including Slot & Dot blot” Submitted to: Dr Khurram Ashfaq Submitted by: Umm-e- Aiman 2017-ag-1435 Department of Pathology

2. Blotting techniques

3. Definition Blotting is a technique for detection of DNA, RNA and Proteins by transferring them on a filter paper after process of gel electrophoresis

4. Types of blotting techniques

5. Blotting sheets Also called whatmman paper World’s most widely used filter paper

6. Southern blotting A procedure to identify specific sequences of DNA through transfer of DNA from agrose gel to filter paper for detection through DNA probe( cDNA ).

7. Northern blotting It is a technique to study gene expression through detection of RNA in samples.

8. Western blotting Proteins are separated by gel electrophoresis and transferred to membrane sheet

9. Dot and slot blot Simplification of western blotting and time saving Proteins not separated through gel electrophoresis Sample is applied on a membrane in a single spot Difference : In dot blotting nucleic acids are blotted as circular blots In slot blotting they are blotted into rectangular slots

10. Steps involved Isolation of DNA/RNA/Protein Loading of sample on agarose gel Blotting on filter paper Labelling with probe Washing to remove unbound probe Detection by autoradiogram

11. Applications Diagnostics Forensics The confirmatory HIV test by detection of antibodies The definitive test for BSE in cow