Team:BIOTEC Dresden/Notebook1-3

From 2009.igem.org

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'''26th August:'''
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1. A PCR reaction has been set up with pR6K-CmR and pR6K-KanR as templates with Kan-R/Kan-F and CmR/CmF primer pairs.
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2. The PCR product has been run on a agarose gel to check quality and later the concentrations are also measured with Nanodrop.
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3. Electroporation of plasmid pR6K-CmR has been done with Pirate competent cells and eventually incubated on LB-Cm15 plates to check for colonies.
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4. Inoculatiuon of pRhaFlp number 6 has been done in LB-Amp and of ptetFlp in LB-Hyg and incubated overnight at 37 degrees shaking.
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'''27th August:'''
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1. Microfluidic templates have been prepared following a standard protocol.
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2. A Red/ET recombination induction step has been carried out for the plasmid pRhaFlp with anhydrotetracycline anf pTetFlp with L-rhamnose and a electroporation has been done at 1350Volts, followed by recovery of cells and plating and incubating overnight for colonies.
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'''28th August:'''
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1. Colonies from pTetFlp-KanR plates are picked and re-inoculated in LB-Kan15 plates.
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2. The GB05 strain is tested for growth in Trimethoprin LB-medium.
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'''29th August:'''
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1. OD measurements of wild type GB05cells and DhfR cells has been done by a spectrophotometer.
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2. Minipreps of pTetFlp-kanR has been done.
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'''31st August:'''
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1. The concentrations of the minipreps of pTetFlp-KanR has been measured using nanodrop.
 +
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2. A restriction digest of the minipreps of pTetFlp-KanR and parental plasmid pRedFlp has been done with PstI.
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3. The digests are run on a 1% agarose gel  and the uncut dna on a 0.9% gel.
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{{:Team:BIOTEC_Dresden/NewTemplateEnd}}

Latest revision as of 02:18, 22 October 2009

26th August:

1. A PCR reaction has been set up with pR6K-CmR and pR6K-KanR as templates with Kan-R/Kan-F and CmR/CmF primer pairs.

2. The PCR product has been run on a agarose gel to check quality and later the concentrations are also measured with Nanodrop.

3. Electroporation of plasmid pR6K-CmR has been done with Pirate competent cells and eventually incubated on LB-Cm15 plates to check for colonies.

4. Inoculatiuon of pRhaFlp number 6 has been done in LB-Amp and of ptetFlp in LB-Hyg and incubated overnight at 37 degrees shaking.

27th August:

1. Microfluidic templates have been prepared following a standard protocol.

2. A Red/ET recombination induction step has been carried out for the plasmid pRhaFlp with anhydrotetracycline anf pTetFlp with L-rhamnose and a electroporation has been done at 1350Volts, followed by recovery of cells and plating and incubating overnight for colonies.

28th August:

1. Colonies from pTetFlp-KanR plates are picked and re-inoculated in LB-Kan15 plates.

2. The GB05 strain is tested for growth in Trimethoprin LB-medium.

29th August:

1. OD measurements of wild type GB05cells and DhfR cells has been done by a spectrophotometer.

2. Minipreps of pTetFlp-kanR has been done.

31st August:

1. The concentrations of the minipreps of pTetFlp-KanR has been measured using nanodrop.

2. A restriction digest of the minipreps of pTetFlp-KanR and parental plasmid pRedFlp has been done with PstI.

3. The digests are run on a 1% agarose gel and the uncut dna on a 0.9% gel.

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