Download and read the SDS-PAGE and Coomassie Stain Protocols. Remember to read through the entire protocol and ask questions about anything you don’t understand before getting started. Reminder: You must submit this assignment before you can continue in the course.
Coomassie Stain Protocol Accessible
You will need to pour your own 15% SDS-PAGE gel, and use it to determine the size of a mystery protein.
Prepare the mystery protein for SDS-PAGE by combining 40 uL of mystery protein + 5 uL of 10x loading dye + 5 uL of 1 M DTT. Mix the resulting solution, and heat it at 95 degrees C for 5 minutes in a heat block. (Alternatively, use PCR tubes and the thermal cycler to heat-treat the samples.) Load 3 lanes of the mystery protein, each 15 uL, to get practice loading these gels. Also load 5 uL of the molecular weight ladder/marker.
Once you have successfully completed the experiment, you will need to submit a figure of your results that includes:
This figure can be made in PowerPoint, and submitted as an image or PDF.
You can sign up for a lab time here: https://biotech.youcanbook.me/
Good luck!