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From 2010.igem.org

9/22/10

Today we have growth from electroporation done yesterday we are going to test by hitting with (AHL) the 20 mL grew not 5 mL sample 1) Scrap bacteria from transformed plate

2) Set up 3) Cuvettes

1 with 200 mL of bacteria + 800 mL LB – Lennox broth

2 with 200 mL of bacteria + 800 mL of A.tumafaciens superant

3 with 200 mL of bacteria + 800 mL of E – Coli Superant

4with 200 mL LB broth + 800 mL of LB

5 with 200 mL LB broth + 800 mL of A tumafaciens

6 with 200 mL LB broth + 800 mL of E – Coli Superant

change 500 mL cells either Agro-T9002 for sample A,B,C or Agro for D,E,F and 500 mL Sup + 200 mL SOB

The only one that should glow should be (C)/(3) E – Coli w/transformed part in LB For Supernate of A.Tumafaciens we will use suspended T9002 in agrobacteria prof T. set up yesterday as our 200 mL spl. - Today I will be electroporating part #’s I732094 (Lacz) – GFP, and part # F2620 (LuxR) into E – Coli Electrocomp cells.

DG pulled the DNA parts from the Bio Brick and placed into 50 mL Electro comp E – Coli cells and 5 ml of DNA spls. now we will electroporate using protocol from pg 15 We recovered cells shooting 900 mL into Cuvette then drawing it off with electroplated cells.

Placed at room temp to grow for 1 hr then place and suspend in LB/Amp (10 mL)

- I took our streaked to try to get an Isolated Colony Streaking and Heating between each pass. then placed into incubator.

- Results after letting incubate for 2 days we have found that the incubator was set to too low of a temp we raised the temp to 37 degrees C from 26 degrees C we will see where we are at on Tuesday.