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

  1. Add experimental record here. Include what, how, and why...

Data

  • Add data and results here...

Notes

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


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.




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