User:University of Washington/27 June 2008

Yeast Shuttle Plasmid
- 1.5 mL 15% glycerol stocks of the overnight BY4741 yeast growth cultures were made by combining 563 uL of 40% glycerol and 937 uL yeast culture, then stored at -80 degrees Celsius.

Conjugation
Mini Prep ·R0062 (LuxR promoter)

Glycerol ·""

Sequencing ·""

UW Glycerol Stocks

LuxR from AraC and TetR
- Made glycerol stock

- Did mini Prep

- Submitted DNA for sequencing reaction and analysis (only for AraC)

LuxR from pLac
- I0462 was again transformed into MG strain again, and plated on AMP.

BioBrick Promoter Measurements
- A strain of TOP10 E. coli was obtained and thawed on ice, along with a strain which had been previously transformed (at MIT) with the fully-assembled reference promoter-RBS-GFP gene-backbone plasmid (I20260).

- 40 uL aliquots of the untransformed cells were transferred into five different tubes. A 40 uL aliquot of the already-transformed cells was similarly transferred into its own tube.

- 2 uL each of plasmid solutions I20260, J23150, J23151, and J23102 were added to four separate aliquots of TOP10 cells.

- The TOP10-pDNA mixtures, the pretransformed TOP10 cells, and the remaining negative control with only untransformed TOP10 cells were incubated on ice for 5 minutes, then incubated in a water bath at 42 degrees Celsius for 30 seconds, then transferred back to ice.

- 250 uL of SOC media was added to each of the TOP10 tubes, then incubated at 37 degrees Celsius on a rotator for 1 hour.

- 100 uL of the each of the transformed cells were placed on Kanamycin plates, then left at room temperature over the weekend to allow for slow growth.

Team:University_of_Washington/Notebook