Team:Newcastle/7 September 2010


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First transformation of 'Bacillius subtilis 168' containing Pmutin4 with Prrnb-GFP containing YneA


The aim of the experiment is to perform insert the plasmid Prrnb-GFP containing YneA which have been ligated eariler into the chromosome of 'Bacillus subtilis 168'. 'B. subtilis' containing the intergated vector will be resistance to both the antibiotic chloramphenicol and streptomycin, therefore those that have successful intergated will be selected with agar plates that contain these antibiotic. The second step will be to identify those colones that have the plasmid intergated at the corerct position in the chromosome, which is the amylase locus. Thus those that have intergardted at the wrong position will not be able to break down starch, which can be tested with the iodine test.

Materials and Protocol

Please refer to: Transformation of Bacillus subtilis Note: Overnigth culture of 'B. subtilis 168' in MM competence medium was done the day before and the iodine test was performed the day after.


Subtilin Immunity BioBrick


The aim of this experiment is to perform minipreps of the 16 overnight cultures that were left overnight from yesterday.


The Qiagen Miniprep protocol was used followed by the NanoDrop Spectrophotometer protocol for each of the 16 cultures.

Results, Discussion and Conclusion

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