User:University of Washington/26 June 2008

Yeast Shuttle Plasmid
- Yeast growth media YEPD was obtained.

- Yeast transforming media 50% PEG, 10x LiAc, and 10x TE were obtained.

- Isolated BY4741 yeast colonies were inoculated in 5 mL of YEPD media and incubated overnight at 30 degrees Celsius in a rotator.

Conjugation Plasmid
Mini Preps ·C600+RP4 construct

Glycerol Stocks ·C600+RP4 ·S17-1

UW Glycerol Stocks

LuxR from AraC and TetR
- Got 1 colony for TetR, 24 for AraC

- Grew culture for Glycerol Stock and Miniprep

LuxR from pLac
- Got 13 colonies for R0010, and 20 colonies for J04430. I0462 failed to grow.

- Grew cultures of R0010 and J04430 for glycerol stock. I0462 transformed e.coli sat out overnight in non-selective media but were plated again on Amp anyway to see if anything would grow.

- Four colonies from the J04430 plate were selected, and re-plated on Amp agar for later use.

BioBrick Promoter Measurements
- Spotted filter paper sections were removed for the GFP reporter gene contained in a plasmid (pSB1A2-E0240), the standard backbone plasmid containing the ccdb toxin (pSB3K3-P1010), the fully-assembled standard reference promoter-RBS-GFP gene-backbone plasmid (I20260), and three different preassembled promoter test constructs with (hopefully) variable transcriptional strengths (J23150, J23151, J23102).

- Filter paper sections were soaked in 5 uL of deionized water for one hour, then centrifuged at 14,000 RPM for 3 minutes.

- The centrifuged plasmids were then stored at 4 degrees Celsius until arrival of the TOP10 E. coli strain.

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