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
|| Plasmid Backbone
||mCherry (mRFP1), works better then original registry part
||2011 Plate 1
||mRFP1 yeast optimized, K/K resistance
||2011 Plate 3
||Used in E. coli
||E. chromi (Red)
||E. chromi (Violet)
||No DNA (Water)
||No DNA (Water)
- Six 1.5 mL tubes
- 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.
- Agar plates were allowed to warm in warm room while other procedures took place.
- They were placed lid down slightly opened.
- DH5α Turbo was taken from -80° freezer and placed on ice to thaw.
- 1.5 mL tubes were labeled 1-6.
- Sample DNA and 10 μL H2O was added to the tubes.
- BioBrick well was located and marked.
- H2O was added to well then exstracted.
- contents were added to tube.
- DH5α Turbo was added to tubes.
- DH5α Turbo is to be kept on ice the entire time.
- Tubes were immediately put on ice for 5 minutes.
- Agar Plates were labeled
- Lab protocol dictated they were to be labeled as follows: Resistance, Strain, Label within experiment, Name of DNA, Initials, Date.
- Contents of tubes were pipetted onto the appropriate plate.
- Contents were spread around plate via 20-25 sterile glass beads.
- 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.
The plates are labeled to indicate what resistance they contain. In this lab a red line indicates Ampicillin and a blur line indicates Kanamycin