IGEM:IMPERIAL/2007/Notebook/2007-8-30: Difference between revisions
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'''Construct - pTet-GFP [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522]'''<br> | '''Construct - pTet-GFP [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522]'''<br> | ||
'''Temperatures - 37 °C and 20°C''' | '''Temperatures - 37 °C and 20°C''' | ||
*For each temperature two experiments carried out at staggered time points to minimize the time points not measured over night. | |||
* | * |
Revision as of 15:43, 30 August 2007
<calendar> name=iGEM:IMPERIAL/2007/Notebook date=2007/09/24 view=threemonths format=%name/%year-%month-%day weekstart=7 </calendar>
Construction of pT7-GFP
- Insert re-digest was run on 1% agarose gel
- 1. 30 μl insert re-digest
- 2. 5 μl uncut insert
- 3. 2 μl 1 kb DNA ladder
Protocols can be found at Protocols can be found at Electrophoresis in the general protocols page
- PCR purified vector digest
- Gel extract purified insert re-digest
Protocols can be found at Protocols can be found at DNA Extaction/Purification in the general protocols page
- Purified vector and insert were run on 1% agarose gel
- 1. 6 μl purified insert
- 2. 3 μl purified vector
- 3. 2 μl 1 kb DNA ladder
Protocols can be found at Protocols can be found at Electrophoresis in the general protocols page
Pictures to be put up soon
- Ligated vector and insert at 14°C overnight
- 7 μl purified insert
- 1 μl purified vector
- 1 μl T4 ligase
- 1 μl T4 ligation buffer
- Set a negative control using ddH20 instead of insert
Cell by date
Operating Temperature Range
Construct - pTet-GFP BBa_I13522
Temperatures - 37 °C and 20°C
- For each temperature two experiments carried out at staggered time points to minimize the time points not measured over night.