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Methods:UV Sensitivity Test

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Contents

Purpose

This test in usually used to test whether a strain is rec-. Rec- strains cannot repair double stranded break due to UV exposure and therefore die.

Materials Needed

  • UV transilluminator
  • piece of cardboard or other opaque material
  • control strains- one rec- and one rec+

Protocol

Before you begin you may need to determine the amount of UV exposure needed to see a difference in growth. UV transilluminators will vary.

Day 1

  • Streak candidates out for single colonies

Day 2

  • Streak candidates in a straight line from 1 end of the plate to the other along with the controls
  • Divide the plate into 4 sections
  • Place the cardboard on the transilluminator
  • Turn the plate over and shield 3/4 of the plate from UV exposure for 10 seconds
  • Move the plate so that only 1/2 of it is shielded for another 10 seconds
  • Move the plate so that only 1/4 of it is shielded for another 10 seconds
  • Turn off the UV
  • Incubate the plate at 37C overnight


Day 3

  • Evaluate growth compared to the controls

[1]

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References

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  1. Miller, JH (1972) Experiments in Molecular Genetics (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY)