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Team:Warsaw/Calendar-Main/3 July 2009
From 2009.igem.org
(Difference between revisions)
| Line 5: | Line 5: | ||
<h4>Marcin</h4> | <h4>Marcin</h4> | ||
<br/> | <br/> | ||
| - | <p> | + | <p>Task 1:</P> |
<ul> | <ul> | ||
| - | <li>Restriction digest of p53 coding sequence obtained from PCR reaction</li> | + | <li>Restriction digest of p53 coding sequence obtained from previously performed PCR reaction</li> |
</ul> | </ul> | ||
<br/> | <br/> | ||
<p>Methods:</p> | <p>Methods:</p> | ||
<ul> | <ul> | ||
| - | <li><p>Reaction mixture composition:</p>15 ul PCR product (DNA concentration about 14.5 ng/ul) | + | <li><p>Reaction mixture composition:</p> |
| - | (Fermentas) | + | <pre>15 ul PCR product (DNA concentration about 14.5 ng/ul) |
| + | 5 ul Tango Buffer (Fermentas) | ||
| + | 1 ul XbaI (Fermentas) | ||
| + | 29 ul MQ water</pre> | ||
</li> | </li> | ||
</ul> | </ul> | ||
| Line 26: | Line 29: | ||
<html> | <html> | ||
<br/> | <br/> | ||
| - | <p> | + | <p>Task 2:</P> |
<ul> | <ul> | ||
<li>ligation of p53 coding sequence with pKS plasmid digested by XbaI and SmaI</li> | <li>ligation of p53 coding sequence with pKS plasmid digested by XbaI and SmaI</li> | ||
| Line 33: | Line 36: | ||
<p>Methods:</p> | <p>Methods:</p> | ||
<ul> | <ul> | ||
| - | <li><p>Ligation mixture composition:</p>15 ul digested p53 | + | <li><p>Ligation mixture composition:</p> |
| - | (Fermentas) | + | <pre>15 ul digested p53 |
| + | 2 ul Tango Buffer (Fermentas) | ||
| + | 1 digested pKS | ||
| + | 0.5 ul ligase T4 | ||
| + | 2 ul 10 uM ATP</pre> | ||
</li> | </li> | ||
</ul> | </ul> | ||
<ul> | <ul> | ||
| - | <li><p>Reaction | + | <li><p>Reaction was carried out about 18 h in 16°C</p> |
</li> | </li> | ||
</html> | </html> | ||
Latest revision as of 11:54, 18 September 2009
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Cloning of p53 coding sequence
Marcin
Task 1:
- Restriction digest of p53 coding sequence obtained from previously performed PCR reaction
Methods:
Reaction mixture composition:
15 ul PCR product (DNA concentration about 14.5 ng/ul) 5 ul Tango Buffer (Fermentas) 1 ul XbaI (Fermentas) 29 ul MQ water
- digest program:
digest
3h 37°C
15 min 80°C
~4°C
Task 2:
- ligation of p53 coding sequence with pKS plasmid digested by XbaI and SmaI
Methods:
Ligation mixture composition:
15 ul digested p53 2 ul Tango Buffer (Fermentas) 1 digested pKS 0.5 ul ligase T4 2 ul 10 uM ATP
Reaction was carried out about 18 h in 16°C
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