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Step-by-Step Guide to Electrophoresis Protocol for Agar Gel Separation

Understand the process and materials needed for agar gel electrophoresis, step-by-step instructions, observation, and conclusion. Electrophoresis separates components based on size and charge. Analyze results and draw conclusions. Ideal for students and researchers. Presented by Jose Manuel Bocara.

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Step-by-Step Guide to Electrophoresis Protocol for Agar Gel Separation

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  1. PROTOCOL FORAGAR ELECTROPHORESIS JoseManuelBocara 

  2. MATERIAL REQUIREDFOR THE PROCESS

  3. MATERIAL NEEDEDFOR EACH PAIR OF STUDENTS

  4. PROCESSESOF ELABORATION

  5. Prepare the TAE 1x solution Dilute 10 ml of TAE (concentration 50x) in 490 ml of destilled water

  6. Prepare yourelectrophoresisbox • Put the comb with 6 wells in the slot. • Place your box on the black paper

  7. Prepare theAgar gel at 1% • Weigh out 1 g of agar powder. • Mix it with 100 ml of TAE 1x. • Heat it in the microwave or on a hotplate, until the solution becomes transparent 1 2 3

  8. ADIAGRAM Not identics Identics

  9. Load thesamples Cut off the gel that has gone over the litle wells using a ruler. Put carbon paper over each end of the box. Pour TAE 1x solution over the gel to cover it. Remove the comb slowly and in a vertical position. Get the syringe and put  a 10 ul tip on the end. Take out spproximately 10 ul of the first solution (to the second line marked on the tip). Put the samples in the first well (or as was decided in point 4). Ensure your hand doesn't shake. Load the other samples into the other wells and do not forget to change your tip each time

  10. Load of Samples images

  11. RuntheGel

  12. Run theGelimages

  13. Observation S2 EC S3 S1 The samples were composed of different colours, each colour moves at a different specific speed in the gel, attracted by the positive pole.

  14. Result Place your box on white paper so you can see the result more clearly

  15. Conclusion • Electrophoresis is a technique that allows us to separate components by size • We can see that some of the samples have 2 components and others have 3. • The yellow component is the smallest and is the most negatively charged as it moves the furthest through the gel towards the positive anode. • The blue components is the largest and has the least negative charge as it moves the least through the gel. Conclusion: Sample S3 has the same components as the crime Scene (EC) sample. S3 S2 EC S1

  16. ENDOFPROTOCOL JoseManuelBocara

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