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Contents 1 rocF Gibson Cloning 1.1 Discussion 2 Subtilin Immunity BioBrick 2.1 Aims 2.2 Materials and protocol 2.3 Results 2.4 Discussion 2.5 Conclusion

rocF Gibson Cloning

Discussion

Today we have colonies from our transformation yesterday. Overnight cultures will be done for DNA extraction tomorrow.

Subtilin Immunity BioBrick

Aims

The aims for today are to perform gel electrophoresis again, and then gel extraction.

Materials and protocol

Please refer to the gel electrophoresis and the gel extraction protocols.

Results

Discussion

We found that the PCR tube containing the spaIFEG gene did not have any band, even though another gel electrophoresis was performed (at 63°C). We had decided to make 5 different PCR tubes with different Tms: 46°C, 51°C, 56°C, 61°C and 66°C. The 6th PCr tube is used as a control, which contains the ara primers - the Tm for this is 59°C, with the extention time of 15 seconds.

We have cut the Plasmid Vector, Promoter & RBS and Double terminator gel bands, following the gel extraction protocol.

Conclusion

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