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.

Results and Conclusion

Please refer to results in tomorrow's lab book.

Transformation

Aim

To transform pGFPrrnB and yneA into E. coli.

Materials and Protocol

Please refer to transformation of E. coli.

Results and Conclusion

Please refer to tomorrow's lab book.

PCR

Aim

To amplify the DNA pSB1C3 using RocF primer.

Materials and Protocol

Please refer to PCR.

Conclusion

Continue with PCR purification.

PCR Purification

Aim

To obtain pure DNA.

Materials and Protocol

Please refer to PCR purification.

rocF and Subtilin immunity

Gel extraction

PCR