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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-=Transformation of 'Bacillius subtilis 168' with Prrnb-GFP containing YneA=
+=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.
-==Aim==
-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: [[Team:Newcastle/Gel electrophoresis| Gel electrophoresis]].
-==Result==
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