IGEM:metu/2009/Notebook/wound dressing/2009/10/05

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Image: Wound dressing logo.jpg

05.10.2009

1. OD measurement (600 nm)

10:30 -- Lard1 : 1,488
         TLiA  : 1,369
         Lard 1 + ABC : 1,463
         TLiA + ABC   : 1,306
13:00 -- Lard1 : 1,169
         TLiA  : 1,060
         Lard 1 + ABC : 1,445
         TLiA + ABC   : 1,050
15:25 -- Lard1 : 1,196
         TLiA  : 0,667
         Lard 1 + ABC : 1,294
         TLiA + ABC   : 1,128
19:45 -- Lard1 : 1,589
         TLiA  : 1,448
         Lard 1 + ABC : 1,524
         TLiA + ABC   : 1,477

2. Preparation of Tris-HCL ( 50mM and pH = 8.5)

3. We found p-nitrophenyl palmitate (pNPP). We will use pNPP while measuring lipase activity.

10 mM pNPP dissolved in acetonitrile was mixed with ethanol and 50 mM Tris-HCl (pH 8.5) to a final ratio of acetonitrile:ethanol:Tris-HCl of 1:4:95 (v/v/v).

4750 μl-- Tris- HCL
200 μl-- EtOH
50 μl-- Acetonitrile

4. We centrifugated our substances and took supernatants.

The reaction will start by adding 50 μl of culture supernatant to 200 μl of reaction mixture.

(Group 2)

5. Changing the strategy about oxygen promoter.Reshaping the strategy accourding to following two papers;

  • Cloning and Expression of the Vitreoscilla Hemoglobin Gene in Enterobacter Aerogenes Effect on Cell Growth and Oxygen Uptake1
  • Degredation of Benzene, Toluene and Xylene by Pseudomonas aeruginosa Engineered with the Vitreoscilla Hemoglobin Gene.

Discussing these two papers and creating a new procedure. Asking the Dr. Geçkil about sending the E.coli strains containing Hemoglobin Gene and oxygen promoter.

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