User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2011/12/13

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Objective

Checking sequences for the GFP mutants today. The PCR was previously run on the wild-type to mutate the aspartate at position 1 to a cysteine. We tried this in class previously, but the primers that I ordered had no mutations in them. New primers were ordered and PCR was run with the following conditions: two reaction vials were started

Vial 1

  • 1uL of WT GFP
  • 1ul of forward primer
  • 1ul dNTP mix
  • 2.5ul reaction buffer
  • 1ul of Pfu Turbo
  • 18.5uL water

Vial 2

  • 1uL of WT GFP
  • 1ul of reverse primer
  • 1ul dNTP mix
  • 2.5ul reaction buffer
  • 1ul of Pfu Turbo
  • 18.5uL water

Each reaction was allowed to proceed for 10 cycles. After 10 cycles the two reaction containers were mixed.

Description

The sequence result can be found here

Data

  • Add data and results here...

Notes

This area is for any observations or conclusions that you would like to note.


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