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

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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.