Team:Newcastle/20 August 2010

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Contents

yneA

Gel Extraction

Aims

To purify the PCR products of digested yneA, pGFPrrnB and pSB1C3 from yesterday, as well as the promoter and double terminator for the rocF BioBrick.

Materials and Protocol

Please refer to gel extraction and nanodrop.

Results

Results of NanoDrop from gel extraction:

DNA Concentration (ng/µl)
yneA 8.5
pGFPrrnB 15.3
pSB1C3 6.4

Discussion

The concentration of yneA, pGFPrrnB and pSB1C3 are not very high.

Conclusion

We continue using these products for ligation.

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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