Team:UCL/Science/Experiment

1. Experiment 01: Extraction of useful BioBrick plasmids from iGEM 2014 Distribution Kit
2. Experiment 02: Transforming E. coli with Azo-reductase plasmids
3. Experiment 03: Diagnostic digest of azo-reductase plasmids
4. Experiment 04: Creation of azo-reductase BioBrick parts from plasmids
5. Experiment 05: Diagnostic digest of azo-reductase BioBrick parts
6. Experiment 06: Assembling azo-reductase BioBrick Device(s)
7. Experiment 07: Characterisation of azo-reductase BioBrick devices

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Experiment 1: Extraction of useful BioBrick plasmids from iGEM 2014 Distribution Kit - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

Background

Content

Results

Content Here

Conclusion

Content Here

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Experiment 2: Transforming E. coli with Azo-reductase plasmids - Date: [10/07/2014]

Protocols
competent cells transformation miniprep digest gel

Background

Performed transformation of the following BioBricks using non-commercial competent cells:

1. BBa_J23119
2. BBa_R0010
3. BBa_R0011
4. BBa_K206000
5. BBa_B0034
6. BBa_B0012
7. BBa_J04450
8. BBa_E2050

Performed transformations of Lisbon plasmids using NEB competent cells (c2987):

1. pPAzoR - P. putida Azo reductase
2. pCotA - Laccase
3. pPpDyP - P.Putida dye decolourizing peroxidase
4. pBsDyP - B. subtilis dye decolourizing peroxidase
5. p1B6 - Azoreductase 1B6 (mutated from AzoR)

Results

Lisbon plasmid transformation using NEB competent cells successfully grew when plated with the appropriate antibiotic

BioBrick plasmid transformations were successful, however the colonies had grown as a lawn making it difficult to pick a single colony.

Conclusion

Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks

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Experiment 3: Diagnostic digest of azo-reductase plasmids - Date: [10/07/2014]

Protocols
competent cells transformation miniprep digest gel

Background

Set up double digest as followed CHECK ENZYMES

Lane	Gene	DNA conc (ng/l)	*Volume (ul)	H2O (ul)
1	AzoR	48	5.21	1.79
2	AzoR1B6	68	3.67	3.33
3	CotA	103	2.42	4.58
4	BsDyP	51	4.91	2.09
5	PyDyp	55	4.54	2.46

Performed transformation of the following BioBricks using non-commercial competent cells:

1. BBa_J23119
2. BBa_R0010
3. BBa_R0011
4. BBa_K206000
5. BBa_B0034
6. BBa_B0012
7. BBa_J04450
8. BBa_E2050

Performed transformations of Lisbon plasmids using NEB competent cells (c2987):

1. pPAzoR - P. putida Azo reductase
2. pCotA - Laccase
3. pPpDyP - P.Putida dye decolourizing peroxidase
4. pBsDyP - B. subtilis dye decolourizing peroxidase
5. p1B6 - Azoreductase 1B6 (mutated from AzoR)

Results

Lisbon plasmid transformation using NEB competent cells successfully grew when plated with the appropriate antibiotic

BioBrick plasmid transformations were successful, however the colonies had grown as a lawn making it difficult to pick a single colony.

Conclusion

Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks

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Experiment 4: Creation of azo-reductase BioBrick parts from plasmids - Date: [17/07/14]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Experiment 5: Diagnostic digest of azo-reductase BioBrick parts - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Experiment 6: Assembling azo-reductase BioBrick Device(s) - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Experiment 7: Characterisation of azo-reductase BioBrick devices - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada