Team:Warsaw/Calendar-Main/4 June 2008

From 2008.igem.org

(Difference between revisions)
Line 3: Line 3:
<html><h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
<html><h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
<h4>Michał K.:</h4><ol>
<h4>Michał K.:</h4><ol>
-
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of DNA fragment  (AIDT7)with HindIII and SalI (2x Tango buffer).</li>
+
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of DNA fragment  (AIDT7) with HindIII and SalI (2x Tango buffer).</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of pMPM-T5 + AID with HindIII and SalI(2x Tango buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> (CIAP).</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of pMPM-T5 + AID with HindIII and SalI(2x Tango buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> (CIAP).</li>

Revision as of 18:37, 5 October 2008

Gallery Bricks Notebook Team Project Home


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

 


Preparation of pMPMT5-AID+AIDT7 construct

Michał K.:

  1. Digest of DNA fragment (AIDT7) with HindIII and SalI (2x Tango buffer).
  2. Digest of pMPM-T5 + AID with HindIII and SalI(2x Tango buffer), dephosphorylation (CIAP).
  3. Clean-up of reaction products.
  4. Ligation of p.4 products (pMPM-T5 + AID transcription fusion with AID-T7 RNA-polymerase translation fusion).
  5. Transformation of E.coli TOP10 with ligation product.
  6. Plating transformants on LB+tetracycline.

Preparation of pMPMT5+T7 construct

Piotr:

  1. Inoculation some separate transformant colonies (liquid LB + tetracycline).