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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> | ||
| Line 15: | Line 15: | ||
<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 | + | <img src="https://static.igem.org/mediawiki/2009/7/7d/PSB_p53_R0080_C0040%2BRBS_gel_out_ocena_18_08_09.png"></br> |
| + | <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 XbaI (Fermentas) | 1 μl XbaI (Fermentas) | ||
0.5 μl SpeI (Fermentas) | 0.5 μl SpeI (Fermentas) | ||
| - | 5 μl Buffer Tango (Fermentas)</pre> | + | 5 μl Buffer Tango (Fermentas)</pre></ul> |
<br/> | <br/> | ||
<p>Task 3:</p> | <p>Task 3:</p> | ||
| Line 40: | Line 41: | ||
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> | ||
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| + | </html> | ||
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| + | <html> | ||
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| + | <h3><div style="text-align: center;">Making of plac-RBS-llo part</div></h3> | ||
| + | <h4>Jarek</h4> | ||
| + | <br /> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Preparation of 20 liquid cultures, from colonies that grew from ligation, in 3 ml of LB+Amp nad incubating them in 37C degree</li> | ||
| + | </ul> | ||
| + | <br /> | ||
| + | <h3><div style="text-align: center;">Cloning switch 1 regulatory parts [ | ||
| + | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012">K177012</a>-PcI.RBS.LacI, | ||
| + | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177033">K177033</a>-PcI.RBS.LacI.PcI.RBS.RFP.terminator, | ||
| + | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K177011">K177011</a>-PLacI.RBS.cI.terminator, | ||
| + | <a href="http://partsregistry.org/Part:BBa_K177038">K177038</a>-PLacI.RBS.cI.terminator.PLacI.RBS.GFP.terminator | ||
| + | ] | ||
| + | into two compatible low copy number plasmids of different antibiotic resistance</div></h3> | ||
| + | <h4>Ania</h4> | ||
| + | <br/> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li> | ||
| + | </li> | ||
| + | </ul> | ||
| + | <br/> | ||
</html> | </html> | ||
Latest revision as of 21:29, 13 September 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)
Making of plac-RBS-llo part
Jarek
Tasks:
- Preparation of 20 liquid cultures, from colonies that grew from ligation, in 3 ml of LB+Amp nad incubating them in 37C degree
Ania
Tasks:
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