Team:Newcastle/20 August 2010

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Contents

Miniprep for yneA, pGFPrrnB and pSB1C3

Aim

The aim of this experiment is to produce stocks of yneA, pGFPrrnB and pSB1C3.

Materials and Protocol

Please refer to miniprep and nanodrop.

Results

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

Table 1: Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.

Discussion

Results from the NanoDrop show concentration values of more than 100 ng/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 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.

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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