Team:Warsaw/Calendar-Main/3 June 2008

(Difference between revisions)

Revision as of 15:09, 24 October 2008


	Attribution analyzer / Interactive list of topics

DNA (PCR products from previous day) gel electrophoresis and isolation of DNA from proper band (20 cycles - 3300 bp). Fig. 1.

Digest of pMPMT5-AID+T7 (transcriptional fusion) with EcoRI and HindIII (2x Tango buffer).
Use of T4 polymerase for blunting (3 hr of incubation).
DNA gel electrophoresis.
Isolation of DNA from proper band (7000 bp).
Ligation of isolated DNA (pMPM-T5 + T7 RNA-polymerase).
Transformation of E.coli TOP10 with ligation product.
Plating transformants on LB + tetracycline.

Fig. 1. PCR results of translational fusion for the pMPMT5-AID+AIDT7 construct. Lane 1 - 25 cycles, lane 2 - 20 cycles.

@@ Line 3: / Line 3: @@
 <html>
-<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
+<h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID%2BAID-T7>pMPMT5-AID+AIDT7</a> construct</h3>
 <h4>Michał K.</h4>
 <p>
@@ Line 18: / Line 18: @@
 <li>DNA gel electrophoresis.</li>
 <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">Isolation of DNA</a> from proper band (7000 bp).</li>
-<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA (pMPM-T5 + T7 RNA-polymerase).</li>
+<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-T7>pMPM-T5 + T7 RNA-polymerase</a>).</li>
 <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with ligation product.</li>
 <li>Plating transformants on LB + tetracycline.</li>