Team:Newcastle/20 August 2010

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{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
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=Transformation of ligated ''yneA'' into pGFPrrnB and pSB1C3=
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==Aims==
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=''yneA''=
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To produce more colonies of the plasmid pGFPrrnB and pSB1C3 containing ''yneA'' with pGFPrrnB and pSB1C3 from[[Team:Newcastle/19_August_2010|yesterday]].
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==Gel Extraction==
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==Materials and Protocol==
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===Aims===
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To purify the PCR products of digested ''yneA'', pGFPrrnB and pSB1C3 from [[Team:Newcastle/19_August_2010|yesterday]], as well as the promoter and double terminator for the ''rocF'' BioBrick.
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===Materials and Protocol===
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Please refer to [[Team:Newcastle/Gel_extraction|gel extraction]] and [[TeamNewcastleNanoDrop_Spectrophotometer|nanodrop]].
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===Results===
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Results of NanoDrop from gel extraction:
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{|border=1
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|-
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! DNA
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! Concentration (ng/µl)
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! ''yneA''
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! 8.5
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! pGFPrrnB
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! 15.3
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! pSB1C3
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! 6.4
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|}
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===Discussion===
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The concentration of ''yneA'', pGFPrrnB and pSB1C3 are not very high.
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===Conclusion===
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We continue using these products for ligation.
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==Miniprep==
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===Aim===
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To produce stocks of ''yneA'', pGFPrrnB and pSB1C3.
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===Materials and Protocol===
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Please refer to [[Team:Newcastle/Qiagen_Minipreps|miniprep]] and [[TeamNewcastleNanoDrop_Spectrophotometer|nanodrop]].
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===Results===
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The table below shows results of concentration (in ng/µl) from NanoDrop:
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{|border=1
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|-
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! Sample no.
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! ''yneA''
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! pGFPrrnB
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! pSB1C3
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!1
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!432.5
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!238.9
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!126.1
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!2
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!347.6
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!230.7
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!107.6
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!3
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!380.2
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!390.9
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!121.5
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!4
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!377.9
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!236.2
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!112.8
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|}
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===Discussion===
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Results from the NanoDrop show concentration values of more than 100ng/µl, which means we have obtained good concentration of DNA from the miniprep.
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===Conclusion===
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We keep the miniprep products as stocks for future use.
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==Transformation of Ligated Products==
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===Aims===
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To produce more colonies of the products of ''yneA'' with pGFPrrnB and pSB1C3 from ligation [[Team:Newcastle/19_August_2010|yesterday]].
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===Materials and Protocol===
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Please refer to [[Team:Newcastle/Transformation_of_E._coli|transformation of E. coli]].
Please refer to [[Team:Newcastle/Transformation_of_E._coli|transformation of E. coli]].
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==Ligation==
 
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===Aims===
 
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To ligate ''yneA'' with pGFPrrnB and ''yneA'' with pSB1C3 (A repeat of [[Team:Newcastle/19_August_2010|yesterday]]).
 
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===Materials and Protocol===
 
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Please refer to [[Team:Newcastle/Ligation|ligation]].
 
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===Results, Discussion and Conclusion===
 
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Please refer to [[Team:Newcastle/23_August_2010|23.08.10]].
 
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==Plating ''Bacillus subtilis'' BFS678==
 
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Has pMutin4 (and thus ''lacI'') integrated into the chromosome...will be useful for characterisation.. will be transforming this strain with our filamentous cell and arginase BioBricks.
 
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 00:33, 28 October 2010

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Transformation of ligated yneA into pGFPrrnB and pSB1C3

Aims

To produce more colonies of the plasmid pGFPrrnB and pSB1C3 containing yneA with pGFPrrnB and pSB1C3 fromyesterday.

Materials and Protocol

Please refer to transformation of E. coli.


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