Team:Newcastle/25 August 2010

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=First transformation of ''Bacillius subtilis'' 168 with Prrnb-GFP containing ''yneA''=
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=Transformation of ''Bacillius subtilis'' 168 with pGFP-rrnB containing ''yneA''=
==Aim==
==Aim==
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To transform ''yneA'' into competent ''B. subtilis''.
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We performed gel electrophoresis on our 12 digested minipreps and found 8 were the correct product. We transformed ''B. subtilis'' 168 with one of the samples (1).
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[[Image:filamentous_in_pgfprrnb.jpg|800px]]
==Materials and Protocol==
==Materials and Protocol==
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Please refer to results in [[Team:Newcastle/26_August_2010|tomorrow]]'s lab book.
Please refer to results in [[Team:Newcastle/26_August_2010|tomorrow]]'s lab book.
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=Restriction digestion and gel extraction linearized pSB1C3=
 
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==Aims==
 
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The aim of this experiment is to digested the plasmid pSB1C3 with the restriction enzyme HindIII to linearize it and and to perform gel extraction to purify it.
 
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==Materials and protocol==
 
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Please refer to the:
 
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*[[Team:Newcastle/Gel_electrophoresis| gel electrophoresis]],
 
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*[[Team:Newcastle/Gel_extraction| gel extraction]] and
 
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*[[TeamNewcastleNanoDrop_Spectrophotometer| NanoDrop spectrophotometer]] protocols.
 
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==Results==
 
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*'''Lane 1''': 1 Kb ladder
 
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*'''Lane 2''': Linearized plasmid pSB1C3
 
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*'''Lane 3''': 1 Kb ladder
 
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There is no gel photograph because we want to keep the exposure of DNA to the UV light to an absolute minimum.
 
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==Discussion==
 
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During gel extraction procedure, we found a bright band of approx
 
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During gel extraction procedure, we found a bright band of approximately 3100 bp size in lane 2 under UV light and we cut the gel and extracted the band.
 
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==Conclusion==
 
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We got linearized plasmid pSB1C3 and we performed gel extraction successfully and the nanodrop protocol showed that we got 12.7 ng/µl concentration of plasmid.
 
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 02:04, 28 October 2010

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Contents

Transformation of Bacillius subtilis 168 with pGFP-rrnB containing yneA

Aim

We performed gel electrophoresis on our 12 digested minipreps and found 8 were the correct product. We transformed B. subtilis 168 with one of the samples (1).

Filamentous in pgfprrnb.jpg

Materials and Protocol

Please refer to transformation of B. subtilis.

Results and Conclusion

Please refer to results in tomorrow's lab book.


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