20.109(F16):Test comet chip assay variables (Day3)
Contents
Introduction
Protocols
Part 1: Examine comet chip loading experiments
In a group discussion with the teaching faculty, you will assess the results of the class data from the comet chip loading experiments. The goal here is to determine which cell number and loading time to use when preparing your comet chip for the tests below.
Be sure to include notes on the discussion and the values for cell number and loading time that you will use in your notebook!
Part 2: Design experiment to optimize comet chip assay
In your experiments from the previous laboratory session, you completed tests to determine the best cell number and loading time to use in the comet chip assay. Today you will examine variables that effect the output. Specifically, you will test conditions that effect the robustness and reproducibility of the comet tails. Remember, the comet tail length is used to measure DNA damage in the comet chip assay.
The two variables that you will test are the length of unwinding time and the length of gel electrophoresis time. Unlike your previous experiment, each team will examine only one variable. Read through your notes regarding the role of unwinding time and electrophoresis time to determine which variable you want to examine today.
Part 3: Complete comet chip assay
Reagents
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