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Team:Warsaw/Calendar-Main/16 July 2009
From 2009.igem.org
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3>Cloning of the mgtc promoter into the pKSII+ plasmid</h3> <h4>Kamil</h4> <br /> <p>Tasks:</p> <ul> <li>Transformant selection</li...) |
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| + | <h3><div style="text-align: center;">Cloning of p53 coding sequence</div></h3> | ||
| + | <h4>Marcin</h4> | ||
| + | <p>Task:</p> | ||
| + | <ul> | ||
| + | <li>Transformation of chemocompetent E. coli strain DH5α</li> | ||
| + | </ul> | ||
| + | <br/> | ||
| + | <p>Methods:</p> | ||
| + | <ul> | ||
| + | <li>Ligation reaction was stopped via thermal inactivation in 80°C for 20 minutes</li> | ||
| + | <li>Detailed protocol of ligation is described <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/5_July_2009">here</a>. The only modification is usage of half volume of ligation mixture prepared 15.07.09</li> | ||
| + | </ul> | ||
| + | <html> | ||
| + | <h3><div style="text-align: center;">Assembly of endosomal detection operon</div></h3> | ||
| + | <h4>Marcin</h4> | ||
| + | <p>Task:</p> | ||
| + | <ul> | ||
| + | <li>Restriction digest of biobricks</li> | ||
| + | </ul> | ||
| + | <br/> | ||
| + | <p><strong>Comment:</strong></p> | ||
| + | <p>Due to obtain set of biobricks which each of them contain RBS and particular coding sequence some of biobricks were digested: | ||
| + | <p> <a href="http://partsregistry.org/Part:BBa_B0032"><span style="color: black">BBa_B0032</a></span></p> | ||
| + | <p> <a href="http://partsregistry.org/Part:BBa_C0040"><span style="color: black">BBa_c0040</a></span></p> | ||
| + | <p> <a href="http://partsregistry.org/Part:BBa_C0051"><span style="color: black">BBa_C0051</a></span></p> | ||
| + | <p> <a href="http://partsregistry.org/Part:BBa_E0032"><span style="color: black">BBa_E0032</a></span></p> | ||
| + | <p>Methods</p></ul> | ||
| + | <li>Digest of BBa_B0032 using SpeI and PstI</li> | ||
| + | <ul><li>Reaction mixture composition: 10 μl purified plasmid DNA product, 0.5 μl SpeI (Fermentas),1 μl PstI (Fermentas), 5 μl Buffer Tango (Fermentas), 34 μl MQ water</li></ul> | ||
| + | <li>Digest of other biobricks using PstI and XbaI</li> | ||
| + | <ul><li>action mixture composition: 10 μl purified plasmid DNA product, 1 μl XbaI (Fermentas),1 μl PstI (Fermentas), 5 μl Buffer Tango (Fermentas), 34 μl MQ water</li></ul> | ||
| + | <li>Both reaction were perform overnight (~12 hours) and both of them were subsequently inactivated via heating in 80deg;C for 20 minutes</li> | ||
| - | + | </html> | |
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{{WarNotebookEnd}} | {{WarNotebookEnd}} | ||
Revision as of 00:18, 21 July 2009
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Cloning of the mgtc promoter into the pKSII+ plasmid
Kamil
Tasks:
- Transformant selection
Methods:
- White colonies were picked up and transferred to a new petri dish.
- Liquid cultures were established along the way for plasmid purification.
- Both the dish and the liquid cultures were incubated at 37°C overnight.
Cloning of p53 coding sequence
Marcin
Task:
- Transformation of chemocompetent E. coli strain DH5α
Methods:
- Ligation reaction was stopped via thermal inactivation in 80°C for 20 minutes
- Detailed protocol of ligation is described here. The only modification is usage of half volume of ligation mixture prepared 15.07.09
Assembly of endosomal detection operon
Marcin
Task:
- Restriction digest of biobricks
Comment:
Due to obtain set of biobricks which each of them contain RBS and particular coding sequence some of biobricks were digested:
Methods
- Reaction mixture composition: 10 μl purified plasmid DNA product, 0.5 μl SpeI (Fermentas),1 μl PstI (Fermentas), 5 μl Buffer Tango (Fermentas), 34 μl MQ water
- action mixture composition: 10 μl purified plasmid DNA product, 1 μl XbaI (Fermentas),1 μl PstI (Fermentas), 5 μl Buffer Tango (Fermentas), 34 μl MQ water
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