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Contents 1 yneA 1.1 Gel Electrophoresis 1.1.1 Aim 1.1.2 Materials and Protocol 1.1.3 Results 1.1.4 Discussion 1.1.5 Conclusion 1.2 Transformation of B. Subtilis 1.2.1 Aim 1.2.2 Materials and Protocol 2 rocF and Subtilin immunity 2.1 Gel extraction 2.2 PCR

yneA

Gel Electrophoresis

Aim

To check if the digestion from yesterday worked.

Materials and Protocol

Please refer to gel electrophoresis for protocol.

Results

The result from gel electrophoresis:

Figure 1 shows the double digest of 12 tubes of pGFPrrnB and yneA.

• Lane 1: Vector only
• Lane 2: Tube 1
• Lane 3: Tube 2
• Lane 4: Tube 3
• Lane 5: Tube 4
• Lane 6: Tube 5
• Lane 7: Tube 6
• Lane 8: Tube 7
• Lane 9: Tube 8
• Lane 10: Tube 9
• Lane 11: Tube 10
• Lane 12: Tube 11
• Lane 13: Tube 12

Discussion

The bands we got from the gel shows that digestion in tubes 1, 2, 4, 10, 11 and 12 worked.

Conclusion

We use the digested products from the six tubes that worked for ligation.

Transformation of B. Subtilis

Aim

To transform yneA into competent B. Subtilis.

Materials and Protocol

Please refer to transformation of B. subtilis.

rocF and Subtilin immunity

Gel extraction

PCR