Team:Warsaw/Calendar-Main/17 June 2008

From 2008.igem.org

(Difference between revisions)
Line 28: Line 28:
<p>Repetition of experiment from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/11_June_2008">11/06/2008.</a></p>
<p>Repetition of experiment from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/11_June_2008">11/06/2008.</a></p>
-
 
-
 
-
<h3>Cloning of protein Z DNA to <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b-OmpA-alpha</a> in place of OmpA</h3><h4>Paweł</h4>
 
-
 
-
<p>Ligation <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformed</a> into <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain and plated on LB + ampicillin.</p>
 

Revision as of 17:33, 24 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 constructs with OmpA protein fusions

Michał K.

  1. PCR on OmpA_linker and linker_alpha with OmpaL_N and AlphaP_XB primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57°C).
  2. PCR on OmpA_linker and linker_omega with OmpaL_N and OmegaP_EPB primers (20 cycles, elongation length 1 min 15 s, annealing temperature 57°C).
  3. Gel electrophoresis of PCR products and gel-out of OmpA_alpha (1000 bp) and OmpA_omega (800 bp).
  4. Electrophoresis to estimate the concentration of isolated DNA.
  5. Overnight digest of purified OmpA_alpha and OmpA_omega DNA with NdeI and BamHI (Tango 2x buffer).
  6. Inoculation of E. coli TOP10 with pACYC177 plasmid into liquid LB + kanamycin.

Blue/white and rifampicin test

Michał L., Ewa

Repetition of experiment from 11/06/2008.

Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/17_June_2008"