Team:Warsaw/Calendar-Main/15 May 2008

From 2008.igem.org

(Difference between revisions)
 
(23 intermediate revisions not shown)
Line 1: Line 1:
{{WarNotebook}}
{{WarNotebook}}
<!-- do not edit above me! -->
<!-- do not edit above me! -->
-
<html>
+
<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a></h3>
 +
<h4>Michał K.</h4>
 +
<p><ol>
 +
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - AID for translational fusion. <br>
-
<p>Michał K.</p>
+
Primers:
-
<p>1. <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical pMPM-T5+AID.</p>
 
-
<p>2. Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digestation</a> of plasmid with HindIII and NcoI (1xTango buffer) - construct control.</p>
+
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDlNrH">AIDlNrH</a>
 +
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDpLinB">AIDpLinB</a>
 +
<br>
-
<p>3. Gel electrophoresis - choice of proper clones.</p>
+
Template DNA: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A-AID</a>
 +
<br>
 +
Annealing temperature: 55&deg;C<br>
 +
Elongation time: 60 s<br>
 +
20 cycles
 +
</li>
 +
 +
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematic PCR).<br>
 +
 +
Primers:
 +
 +
 +
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7lLinkB">T7lLinkB</a>
 +
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7pXbSal">T7pXbSal</a>
 +
<br>
 +
 +
 +
Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a>  genomic DNA<br>
 +
 +
Annealing temperature: 62&deg;C<br>
 +
 +
Elongation time: 4 minutes<br>
 +
 +
20 cycles<br>
 +
</li>
 +
 +
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA-polymerase for transcriptional fusion.<br>
 +
 +
Primers:
 +
 +
 +
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7lRBSHi">T7lRBSHi</a>
 +
<a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7pXbSal">T7pXbSal</a>
 +
<br>
 +
 +
 +
Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a>  genomic DNA<br>
 +
 +
Annealing temperature: 62&deg;C<br>
 +
 +
Elongation time: 4 minutes<br>
 +
 +
20 cycles
 +
</li>
 +
</p>
</html>
</html>
<!-- do not remove this! -->
<!-- do not remove this! -->
{{WarNotebookEnd}}
{{WarNotebookEnd}}

Latest revision as of 13:46, 26 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
Interactive list of topics
next notebook entry

 


Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

  1. PCR - AID for translational fusion.
    Primers: AIDlNrH AIDpLinB
    Template DNA: pTrc99A-AID
    Annealing temperature: 55°C
    Elongation time: 60 s
    20 cycles
  2. PCR - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematic PCR).
    Primers: T7lLinkB T7pXbSal
    Template DNA: E. coli Rosetta genomic DNA
    Annealing temperature: 62°C
    Elongation time: 4 minutes
    20 cycles
  3. PCR - T7 RNA-polymerase for transcriptional fusion.
    Primers: T7lRBSHi T7pXbSal
    Template DNA: E. coli Rosetta genomic DNA
    Annealing temperature: 62°C
    Elongation time: 4 minutes
    20 cycles

Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/15_May_2008"