IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/07/16

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Practice GFP Assay - Does it glow at all?

1) Stat a single colony of 186W from 7.14.09 replate. Started at 11 am

2) reached OD of 0.2 at 3 Pm. Put paraoxan to final concentration of 1 mM

3) dilute 3.6 ml of 100 mM in MeOH with 3.6 ml of dH2O

Final [] of stock is 50 mM in 50% EtOH

-dilute 1ml final [] 20 mM

added 0.4 ml 50 mM stock, 0.6 ml dd H2O

-> final is 20% MeOH 20mM

-put 50 ul 20 mM stock into 950 uL culture (OD = 0.2)

- incubate starting at ~ 4PM. Pick it up at 6 PM (generally, allot 2 hours of incubation time)

- looked under microscope.

                               Result = NO GFP IN FIRST TRY

-Replated his - Paraoxan Biosensor colonies. 3-4 colonies/plate.

-put into incubator

- need to replate Kan - Paraoxan plates tomorrow. Can already see lots of cells

July 16, 2009 P2

Transformations of Longtine PCR Products into Yeast

Transformed products into yeast, however accidentally used Uracil plates instead of -HIS plates for products 1-3. Transformations were unsuccessful for 4 and 5. Beads may spread cell too thinly. Pipet shearing may have also been a cause.