Team:Warsaw/Calendar-Main/30 September 2008

From 2008.igem.org

(Difference between revisions)
Line 15: Line 15:
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of isolated plasmids with EcoRI and BcuI (BamHI buffer) <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008#fig1">Fig. 1</a><a name="fig1">. </li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of isolated plasmids with EcoRI and BcuI (BamHI buffer) <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008#fig1">Fig. 1</a><a name="fig1">. </li>
-
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA_omega_deltaA_alpha</a> with BamHI and PstI (BamHI buffer) <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008#fig2">Fig. 1</a><a name="fig2">. </li>
+
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>pACYC177+OmpA_omega_deltaA_alpha</a> with BamHI and PstI (BamHI buffer) <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008#fig2">Fig. 2</a><a name="fig2">. </li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">Dephosphorylation</a> (CIAP) of plasmids.</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">Dephosphorylation</a> (CIAP) of plasmids.</li>
Line 39: Line 39:
<a name="fig1"><https://static.igem.org/mediawiki/2008/0/01/Go2_30_09.jpg" width=300/></a> </a><var><b>Fig. 1. Control EcoRI/BcuI digests of isolated plasmids</b><br>  
<a name="fig1"><https://static.igem.org/mediawiki/2008/0/01/Go2_30_09.jpg" width=300/></a> </a><var><b>Fig. 1. Control EcoRI/BcuI digests of isolated plasmids</b><br>  
1. Marker<br>
1. Marker<br>
-
2-3. digested plasmids  <br></var>
+
2-3. digested plasmids BBa_I739204 <br></var>
-
4. digested plasmid  
+
4. digested plasmid psB2K3
</html>
</html>

Revision as of 20:22, 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

 


Mutagenesis of protein A

Paweł

Results of sequencing: unfortunately all sequences were wild-type.

Preparation of BioBricks

Michał K.

  1. Isolation of pSB2K3 and BBa_I739204 (pACYC177 converted into BioBrick vector) plasmids.
  2. Digest of isolated plasmids with EcoRI and BcuI (BamHI buffer) Fig. 1.
  3. Digest of pACYC177+OmpA_omega_deltaA_alpha with BamHI and PstI (BamHI buffer) Fig. 2.
  4. Dephosphorylation (CIAP) of plasmids.
  5. Gel elctrophoresis and gel-out of proper band: ??????.
  6. Digest of omega_link fragment (PCR from previous day) with EcoRI and BcuI (BamHI buffer)
  7. Digest of OmpA_omega with BglII and PstI (Orange buffer).
  8. Clean-up of above digest reactions.
  9. Overnight digest of purified pLac_OmpA_omega fragment (from previous day) with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.

Piotr

  1. E. coli TOP10 transformation with overnight ligations pSB1A3 + Z(BBa_K103004) and pSB1A3 + OmpA_link).
  2. Plating on LB + ampicillin.
  3. Inoculation of bacteria received from iGEM HQs, carrying pSB2K3 and BBa_I739204 (pACYC177 converted into BioBrick vector) plasmids.
Fig. 1. Control EcoRI/BcuI digests of isolated plasmids
1. Marker
2-3. digested plasmids BBa_I739204
4. digested plasmid psB2K3


Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008"