Team:Newcastle/20 August 2010

From 2010.igem.org

(Difference between revisions)
(Subtilin Immunity and rocF)
(Gel Extraction)
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=''yneA''=
=''yneA''=
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==Gel Extraction==
 
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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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|-
 
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! ''yneA''
 
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! 8.5
 
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|-
 
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! pGFPrrnB
 
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! 15.3
 
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|-
 
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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.
 
==Miniprep==
==Miniprep==

Revision as of 02:42, 26 October 2010

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Contents

yneA

Miniprep

Aim

To produce stocks of yneA, pGFPrrnB and pSB1C3.

Materials and Protocol

Please refer to miniprep and nanodrop.

Results

The table below shows results of concentration (in ng/µl) from NanoDrop:

Sample no. yneA pGFPrrnB pSB1C3
1 432.5 238.9 126.1
2 347.6 230.7 107.6
3 380.2 390.9 121.5
4 377.9 236.2 112.8

Discussion

Results from the NanoDrop show concentration values of more than 100ng/µl, which means we have obtained good concentration of DNA from the miniprep.

Conclusion

We keep the miniprep products as stocks for future use.


Transformation of Ligated Products

Aims

To produce more colonies of the products of yneA with pGFPrrnB and pSB1C3 from ligation yesterday.

Materials and Protocol

Please refer to transformation of E. coli.


Ligation

Aims

To ligate yneA with pGFPrrnB and yneA with pSB1C3 (A repeat of yesterday).

Materials and Protocol

Please refer to ligation.

Results, Discussion and Conclusion

Please refer to 23.08.10.



Plating Bacillus subtilis BFS678

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.


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