Steps to Conduct an SDS PAGE Gel Electrophoresis

What is the correct order for conducting an SDS PAGE gel? Option 1: Pour the resolving gel, pour the stacking gel, load the samples, stain, destain

Correct Order for Conducting an SDS PAGE Gel

The correct order for conducting an SDS PAGE gel is:
  1. Pour the resolving gel
  2. Pour the stacking gel
  3. Load the samples
  4. Stain the gel
  5. Destain to visualize the proteins

Explanation:

SDS Polyacrylamide Gel Electrophoresis (PAGE) is a technique used to separate proteins based on their size. The process involves various steps and requires a specific ordering for successful protein separation.

Step-by-Step Process:

  1. Pour the resolving gel: This step involves creating a matrix with larger pores that allow proteins to move through. It facilitates the separation of proteins based on size.
  2. Pour the stacking gel: The stacking gel, with smaller pores, is layered on top of the resolving gel to concentrate proteins into a single band.
  3. Load the samples: Carefully load the protein sample onto the gel, avoiding sample mixing.
  4. Stain the gel: After the proteins have run through the gel, use a stain like Coomassie blue to visualize the protein bands.
  5. Destain: Finally, destain the gel to remove excess stain, leaving only the protein bands visible for analysis.

Remember, SDS PAGE gels are run vertically, with proteins moving towards the positive electrode and separating based on size.

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