Team:Newcastle/25 August 2010

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{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
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=Gel electrophoresis for single digestion of pSB1C3=
 
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==Aim==
 
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The aim of this experiment is to check if the digestion from [[Team:Newcastle/24_August_2010|yesterday]] worked.
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=Transformation of ''Bacillius subtilis'' 168 with pGFP-rrnB containing ''yneA''=
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==Materials and Protocol==
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==Aim==
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Please refer to [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] for 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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==Results==
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[[Image:filamentous_in_pgfprrnb.jpg|800px]]
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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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=First transformation of ''Bacillius subtilis'' 168 with Prrnb-GFP containing ''yneA''=
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==Aim==
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To transform ''yneA'' into competent ''B. subtilis''.
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==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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