Title: Milestone Lab: Completing an SDS-PAGE
Course: BTEC-2020-301-302-303-F21 > Section: Protein Gels

Module2 Milestone:Completing an SDS-PAGE

Lab

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.

SDS-PAGE protocol

SDS-PAGE Accessible

Coomassie Stain Protocol

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:

- An appropriate title
- A picture of the stained gel
- Labels of what is in each lane
- Labels of the sizes of the ladder bands
- Labels indicating the sizes of the unknown bands

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!