User:Derek J. Sanchez/Notebook/TB Sensor - MS applied project/2012/03/06

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03/06/12

Transformation of Reporter

The transformation of DNA reporters from BioBricks was preformed.
The following table contains the information about the DNA being transformed into the E. Coli.

Sample Label BioBrick part number Origin Well Plasmid Backbone Organism Resistance Description
1 BBa_J06504 From Yue pSB1A2 Bacteria Amp mCherry (mRFP1), works better then original registry part
2 BBa_E2060 2011 Plate 1 12F pSB2K3 Yeast Kam mRFP1 yeast optimized, K/K resistance
3 BBa_K274100 2011 Plate 3 6H pSB1A2 Used in E. coli Amp E. chromi (Red)
4 BBa_J06702 From Yue pSB1T3 E. coli Amp E. chromi (Violet)
5 No DNA (Water) Lab None N/A Amp Control
6 No DNA (Water) Lab None N/A Kam Control


Materials Used

  • Six 1.5 mL tubes
  • Pipettes
  • A bunch of pipette tips
  • 6 Agar plates
    • 4 with 100μg/mL Ampicillin
    • 2 with 50μg/mL Kanamycin
  • DH5α Turbo chemically competent cells 200μL/Tube
    • Each transformation requires 30μL.


Procedure Used

  1. Agar plates were allowed to warm in warm room while other procedures took place.
    They were placed lid down slightly opened.
  2. DH5α Turbo was taken from -80° freezer and placed on ice to thaw.
  3. 1.5 mL tubes were labeled 1-6.
  4. Sample DNA and 10 μL H2O was added to the tubes.
    1. BioBrick well was located and marked.
    2. H2O was added to well then exstracted.
    3. contents were added to tube.
  5. DH5α Turbo was added to tubes.
    DH5α Turbo is to be kept on ice the entire time.
  6. Tubes were immediately put on ice for 5 minutes.
  7. Agar Plates were labeled
    Lab protocol dictated they were to be labeled as follows: Resistance, Strain, Label within experiment, Name of DNA, Initials, Date.
  8. Contents of tubes were pipetted onto the appropriate plate.
  9. Contents were spread around plate via 20-25 sterile glass beads.
  10. Beads then removed and plates put in warm room to incubate, lid side down.
    Standard incubation time approximately 8 hours. This procedure however was done with a few hours in warm room then left over night at room temperature.

Additional Notes

The plates are labeled to indicate what resistance they contain. In this lab a red line indicates Ampicillin and a blur line indicates Kanamycin




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