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Team:Warsaw/Calendar-Main/18 August 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3><div style="text-align: center;">Assembly of endosomal detection operon</div></h3> <h4>Marcin</h4> <br/> <p>Task 1:</p> <ul> <li>...) |
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<p>Task 1:</p> | <p>Task 1:</p> | ||
<ul> | <ul> | ||
| - | <li>Gel-outs of following constructs digested | + | <li>Gel-outs of following constructs digested on 17.08.09:</li> |
<ul><li>p53 CDS on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmid</li> | <ul><li>p53 CDS on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmid</li> | ||
<li><a href="http://partsregistry.org/Part:BBa_R0080"><span style="color: black">BBa_R0080</a></span> on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmid</li> | <li><a href="http://partsregistry.org/Part:BBa_R0080"><span style="color: black">BBa_R0080</a></span> on <a href="http://partsregistry.org/Part:pSB1A3"><span style="color: black">pSB1A3</a></span> plasmid</li> | ||
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<p>Methods:</p><ul> | <p>Methods:</p><ul> | ||
<li>All samples were inactivated via heating in 80 °C for 20 minutes | <li>All samples were inactivated via heating in 80 °C for 20 minutes | ||
| - | <li>All gel-outs were | + | <li>All gel-outs were performed using the EurX gel-out kit according to the manual</a></li></ul> |
<p>Results</p> | <p>Results</p> | ||
| - | <ul><li>All gel-outs (except C0040 with RBS) were successful. One isolation | + | <ul><li>All gel-outs (except C0040 with RBS) were successful. One isolation failed because of very low DNA concentration</ul></li></br> |
<p>Task 2:</p> | <p>Task 2:</p> | ||
<ul> | <ul> | ||
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1 μl T4 ligase (Fermentas) | 1 μl T4 ligase (Fermentas) | ||
</pre> | </pre> | ||
| - | <li>Ligation was | + | <li>Ligation was performed for about 9 hours and it was subsequently thermally inactivated</li> |
</ul> | </ul> | ||
Revision as of 11:47, 19 August 2009
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Assembly of endosomal detection operon
Marcin
Task 1:
- Gel-outs of following constructs digested on 17.08.09:
Methods:
- All samples were inactivated via heating in 80 °C for 20 minutes
- All gel-outs were performed using the EurX gel-out kit according to the manual
Results
- All gel-outs (except C0040 with RBS) were successful. One isolation failed because of very low DNA concentration
Task 2:
- Digest pKSII plasmid with cro CDS
Methods:
- Reaction mixture composition:
45 μl purified plasmid DNA product 1 μl XbaI (Fermentas) 0.5 μl SpeI (Fermentas) 5 μl Buffer Tango (Fermentas)
Task 3:
Methods:
- Reaction mixture composition:
10 μl p53 CDS 7 μl vector 2.3 μl Buffer Tango (Fermentas) 3 μl dNTPs mixture (EurX) 1 μl T4 ligase (Fermentas)
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