Team:Newcastle/25 August 2010

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{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
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=''yneA''=
 
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==Gel Electrophoresis==
 
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===Aim===
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=Transformation of ''Bacillius subtilis'' 168 with pGFP-rrnB containing ''yneA''=
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To check if the digestion from [[Team:Newcastle/24_August_2010|yesterday]] worked.
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==Aim==
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===Materials and Protocol===
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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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Please refer to [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] for protocol.
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[[Image:filamentous_in_pgfprrnb.jpg|800px]]
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===Results===
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==Materials and Protocol==
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The result from gel electrophoresis:
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[[Image:Newcastle 25-08-2010 – Gel 1 (Ethidium Bromide).png|500px|centre]]
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'''Figure 1''' shows the double digest of 12 tubes of pGFPrrnB and ''yneA''.
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*'''Lane 1''': Vector only
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*'''Lane 2''': Tube 1
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*'''Lane 3''': Tube 2
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*'''Lane 4''': Tube 3
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*'''Lane 5''': Tube 4
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*'''Lane 6''': Tube 5
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*'''Lane 7''': Tube 6
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*'''Lane 8''': Tube 7
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*'''Lane 9''': Tube 8
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*'''Lane 10''': Tube 9
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*'''Lane 11''': Tube 10
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*'''Lane 12''': Tube 11
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*'''Lane 13''': Tube 12
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===Discussion===
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The bands we got from the gel shows that digestion in tubes 1, 2, 4, 10, 11 and 12 worked.
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===Conclusion===
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We use the digested products from the six tubes that worked for ligation.
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==Transformation of ''B. Subtilis''==
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===Aim===
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To transform ''yneA'' into competent ''B. Subtilis''.
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===Materials and Protocol===
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Please refer to [[Team:Newcastle/Transformation_of_B._subtilis|transformation of ''B. subtilis'']].
Please refer to [[Team:Newcastle/Transformation_of_B._subtilis|transformation of ''B. subtilis'']].
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===Results and Conclusion===
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==Results and Conclusion==
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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=''rocF'' and Subtilin immunity=
 
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==Gel extraction==
 
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==PCR==
 
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[[Image:Newcastle_25810gel_psb1c3.jpg|500px]]
 
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{{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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