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Team:Newcastle/20 August 2010
From 2010.igem.org
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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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