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Contents 1 pH changing for HEPES 2 Transformation of Bacillius subtilis 168 with pGFP-rrnB containing filamentous cell part 2.1 Aim 2.2 Materials and Protocol

pH changing for HEPES

We prepared 100mM of HEPES (it is a dibasic compound) at pH 7.0 and sent it for autoclaving. N.B. During autoclaving we loose some volume of the solution.

Transformation of Bacillius subtilis 168 with pGFP-rrnB containing filamentous cell part

Aim

The aim of the experiment is to transform Bacillus subtilis 168 with the plasmid pGFP-rrnB containing our filamentous cell part. After transformation the plasmid will integrate into the chromosome at amyE. This integration removes the ability to break down starch, allowing us to select for our desired construct using 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 transformation.