User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2012/06/11

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Objective

Transform PCR and test plasmids into competent cells

Description

  1. Four Samples for transformation
    1. Novagen Test Plasmid into NovaBlue GigaSingles (recently purchased)
    2. Novagen Test Plasmid into M15MA competent cells (recently made) - cells sent from David Tirrell at Caltech
    3. A71M M8S M33S M103S Ascaris Hb into NovaBlue GigaSingles (recently purchased)
    4. WT Ascaris Hb into NovaBlue GigaSingles (recently purchased)

Plates:

  1. 1.875 g LB
  2. 1.5 g Agar
  3. 75mL water
  4. autoclave
  5. 75uL of 100mg/mL ampicillin after mixture had cooled but not set

Transformation protocol

  1. Place plastic culture tubes on ice for 15 minutes
  2. Place DNA for transformation on ice
  3. After DNA has thawed, place cells on ice. (Note - want to minimize the time that the cells are out of freezer and on ice)
  4. Add 1uL of DNA to the appropriately labeled culture tube and let sit in ice for 1 minute
  5. Add cells (30uL, 40uL, or 50uL) to the tube on top of the DNA and gently pipette up and down to mix cells and DNA
  6. Put tube back in ice for 4 minutes
  7. Heat shock at 42C for 80seconds
  8. Add 100uL of SOC media
  9. Shake at 37C for 1 hour
  10. Plate 50uL of culture media

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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