Team:Warsaw/Calendar-Main/19 May 2008

From 2008.igem.org

(Difference between revisions)
Line 2: Line 2:
<!-- do not edit above me! -->
<!-- do not edit above me! -->
<html>
<html>
-
<h4>Piotr:</h4>
+
<p>
 +
<ol>
 +
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - translation fusion: AID + T7 RNA-polymerase<br>
-
<p>1. Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/16_May_2008">16 May</a> PCR's) and DNA <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> from proper bands (600 bp for AID lane and 2700 bp for T7 polymerase lanes).</html>
+
Primers: <br>
 +
 
 +
<ul>
 +
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDlNrH">AIDlNrH</a></li>
 +
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7pXbSal">T7pXbSal</a></li>
 +
</ul>
 +
 
 +
 
 +
 
 +
Template DNA: purified PCR products from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/16_May_2008">16 May</a> - AID and T7 RNA-polymerase for translation fusion <br>
 +
 
 +
Annealing temperature : 73&deg;C <br>
 +
Elongation time: 4 minutes <br>
 +
 
 +
20 cycles <br></li>
 +
<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of proper band (3300 bp).</li></ol></p>
 +
</html>
<html>
<html>

Revision as of 11:12, 5 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
next notebook entry

 


  1. PCR - translation fusion: AID + T7 RNA-polymerase
    Primers:
    Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation fusion
    Annealing temperature : 73°C
    Elongation time: 4 minutes
    20 cycles
  2. Gel electrophoresis and isolation of proper band (3300 bp).

Michał L., Ewa, Marcin:

1. Overnight inoculation and induction:

Top10 + 0,5 mM IPTG

pMPM-AID + Glc + 0,01% L-Ara

pTrc99a-AID + Glc + 0,5 mM IPTG