First transformation of Bacillius subtilis 168 with Prrnb-GFP containing yneA

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.

Restriction digestion and gel extraction linearized pSB1C3

Aims

The aim of this experiment is to digested the plasmid pSB1C3 with the restriction enzyme HindIII to linearize it and and to perform gel extraction to purify it.

Materials and protocol

Please refer to the:

• gel electrophoresis,
• gel extraction and
• NanoDrop spectrophotometer protocols.

Results

• Lane 1: 1 Kb ladder
• Lane 2: Linearized plasmid pSB1C3
• Lane 3: 1 Kb ladder

There is no gel photograph because we want to keep the exposure of DNA to the UV light to an absolute minimum.

Discussion

During gel extraction procedure, we found a bright band of approx During gel extraction procedure, we found a bright band of approximately 3100 bp size in lane 2 under UV light and we cut the gel and extracted the band.

Conclusion

We got linearized plasmid pSB1C3 and we performed gel extraction successfully and the nanodrop protocol showed that we got 12.7 ng/µl concentration of plasmid.