EMSA_Presentationn_techniqqueee (1).pptx

EMSA is a technique used to detect the binding between a nucleic acid (DNA or RNA) and a protein by observing changes in the mobility of the nucleic acid during gel electrophoresis.

1. GOVT HOLKAR SCIENCE COLLEGE , INDORE TOPIC :- Electrophoretic Mobility Shift Assay (EMSA) SUBJECT :- BACTERIAL GENETICS SESSION :- 2024-25 SUBMITTED TO:- ANKITA MAM SUBMITTED BY:- HARSH PANCHAL

2. Electrophoretic Mobility Shift Assay (EMSA) - A technique to study DNA-protein interactions - Also known as Gel Shift Assay - Used widely in molecular biology

3. Introduction to EMSA - Detects protein binding to specific DNA sequences - Based on mobility change during electrophoresis - Non-denaturing gel is used to retain interactions

4. Principle of EMSA - Free DNA migrates faster than DNA-protein complex - DNA-protein complexes show mobility shift - Shift indicates binding between DNA and protein

5. Applications of EMSA - Studying transcription factor binding - Analyzing DNA-protein interaction specificity - Validating promoter and enhancer regions

6. Materials Required - Labeled DNA probe (radioactive or fluorescent) - Protein extract (nuclear or cell lysate) - Non-denaturing polyacrylamide gel

7. Basic Procedure - Incubate labeled DNA with protein extract - Run on a non-denaturing polyacrylamide gel - Visualize DNA-protein complex shift

8. Controls Used - Free probe to show unbound DNA - Competitor DNA for specificity check - Supershift with antibody to identify protein

9. Advantages of EMSA - Highly sensitive to binding events - Works with small DNA quantities - Useful for qualitative analysis

10. Limitations of EMSA - Not quantitative without additional methods - Does not identify unknown proteins directly - Requires optimization for each system

11. Conclusion - EMSA is vital for studying gene regulation - Still widely used despite new techniques - Combines simplicity with powerful insight