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Team:Newcastle/7 September 2010
From 2010.igem.org
(New page: {{Team:Newcastle/mainbanner}} =First transformation of 'Bacillius subtilis 168' containing Pmutin4 with Prrnb-GFP containing YneA= ==Aim== The aim of the experiment is to perform insert ...) |
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=First transformation of 'Bacillius subtilis 168' containing Pmutin4 with Prrnb-GFP containing YneA= | =First transformation of 'Bacillius subtilis 168' containing Pmutin4 with Prrnb-GFP containing YneA= | ||
| - | ==Aim== | + | ===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. | 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== | + | ===Materials and Protocol=== |
Please refer to: [[Team:Newcastle/Transformation of B. subtilis| Transformation of ''Bacillus subtilis'']] | 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. | 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. | ||
| - | ==Result== | + | ===Result=== |
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| + | '''Go back to our main [[Team:Newcastle/notebook| Lab book]] page''' | ||
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Revision as of 09:40, 8 September 2010
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Contents |
First transformation of 'Bacillius subtilis 168' containing Pmutin4 with Prrnb-GFP containing YneA
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: 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.
Result
Go back to our main Lab book page
   
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