Team:SHPH-Shanghai/Result

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1.Result & Analysis

2.Gene Extraction

3.PCR

Result and analysis

(1) Result of the Pre-experiment

In each plate, from left to right, the pH value for each hole is respectively 1,3,5,7.

3 days cultured in neutral medium before adding the acidic medium

4 days cultured in neutral medium before adding the acidic medium

5 days cultured in neutral medium before adding the acidic medium

(2) Analysis of the Pre-experiment

From both the photography of the 96-microwell plate and the value of OD in each hole, we can observe a significant relevance between the level of biofilm degradation and acidic culturing environment. In the graph, biofilms under condition of pH=1 and pH=3 were very thin and hard to observe, which is consistent with the OD value. On the other hand, biofilms under the condition of pH=5 and 7, the degradation hardly took place.

From the perspective of designing the investigating subject, biologically modifying the biofilm living condition to pH=3 would be a proper and achievable aim. For pH=1, it is too hard for any genetically modified bacteria to achieve; and the effect of pH=1 and pH=3 are similar with regard to the degradation level of the biofilm.

Gene Extraction

Bacterial gene extraction: we use the standard protocol included in the extraction kit. Tissue gene extraction: we used the standard protocol included in the extraction kit.

PCR

(1)PCR of S-layer protein

(2)PCR of NAG

(3)Fusion PCR