Team:UCLA/Notebook/Protein Cages/13 July 2015

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Intro: Miniprep Yeates' wt cage cells. Assisted Phil in making reagents. Lab meeting. Prepared protocols for transformation.

Miniprep Used 2mL of 10mL overnight starter culture. Used highest centrifuge setting for all centrifugations. Yield: 108.01 ng/uL 1.98 260/280. stored in -20C boelter protein cage box.

Notes: Remember to put cap on centrifuge. Rcf dorresponds to g's. Switch between using rpm/rcf button. Balance all tubes with 1.5mL tube and water. Not super precise but its ok. The P3 buffer is in the fridge with the glass door.

Solution making technique: Rotate solid reagent bottle to pout out precise amount. Add acid/base to water. If no flat surface, hold bottle with two fingers to level it.

Lab meeting notes: Think of types of people to contact for human practices. Wearable biomaterials. iGEM journal club next Wednesday.

Michael Gel making: Mix agar and TAE buffer. Microwave for a couple minutes. Let it boil for about 10 seconds before stopping microwave. Add cybrsafe. All casting trays/electrophoresis machines in 7731.

Tyler Lee --Wtleeiv 22:46, 14 July 2015 (CDT)

Phillip's notes:

Introduction: Inoculation into the large culture, and protein induction, will be done today.

Procedures:

Inoculation: 2mL of the starter culture was given to Tyler in order to miniprep. The starter culture was taken out at 12:00PM at left on the bench while autoclaving a 2L flask. Upon cooling the flask, 1L of LB and 1mL of 100mg/mL carbenicillin was added to the flask. The remaining 8mL of starter culture was poured into the flask at 1:30PM. Induction: At 3:30PM (2 hours) 0.0477g of IPTG was dissolved in 1mL of LB, and added to the flask for a total concentration of 0.2mM. The OD600 was measured to be 0.165. Because there was no access to an 18C incubator, the flask was left in the 30C incubator.

Observations: OD600 should have been measured before induction, as the ideal OD600 at time of induction should be 0.6. Also, the flask should have been sterilized the day before in order to not have to leave the bacteria culture on the bench.