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1.Directly testing the oxygen promoter in the presence or absence of oxygen.We have prepared 4 set of tubes in the presence or absence of oxygen and we compared the growth of bacterias by using spectrophotometer. OR, testing the hemoglobin activity; we thought that if the gene coding for hemoglobin is working it means that our oxygen promoter is also working.
2. Centrifuge of supernatant + ammonium sulfate, 14000 rpm for 30 minutes.
3. Preparation of potassium phosphate buffer ( pH 6.6)
4. Resuspend the pellet in potassium phosphate buffer, 5ml,0.1 M and pH 6.6
5.Dialyzing them (Lard, Lard + ABC and Only top 10) in a nitrocellulose tube at 4 centigrade degrees in 500 microliter potassium phosphate buffer stirring for overnight.