Latest revision as of 01:53, 28 October 2010

Preparation for pH acclimatisation of Bacillus subtilis 168

We prepared 100mM of HEPES (it is a dibasic compound) at pH 7.0 and sent it for autoclaving.

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-=pH changing for HEPES=
+=Preparation for pH acclimatisation of ''Bacillus subtilis'' 168=
 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.
-[[Image:filamentous_in_pgfprrnb.jpg|800px]]
-==Materials and Protocol==
-Please refer to: [[Team:Newcastle/Transformation of B. subtilis| 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.
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