Team:Warsaw/Calendar-Main/7 July 2008

From 2008.igem.org

(Difference between revisions)
Line 26: Line 26:
</table>
</table>
-
<h4 style="test-align: center">PCR program for linker-A and omega-linker</h4>
+
<thead><b> Results of rifampicin test</b><br></thead>
<table id="result">
<table id="result">

Revision as of 14:28, 11 October 2008

Gallery Bricks Notebook Team Project Home

Results of rifampicin test

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

 


Preparation of constructs with OmpA protein fusions

Piotr

  1. Digest of confirmed clones of pET15b+OmpA_alpha and pET15b+OmpA_omega with NdeI and BamHI (Tango 2x buffer).
  2. Digest of pACYC177 plasmid with NdeI and BamHI (Tango 2x buffer), dephosphorylation with CIAP.
  3. Gel electrophoresis of digested plasmids and gel-out of proper bands (OmpA_alpha- 1000 bp, OmpA_omega - 800 bp).
  4. Electrophoresis of gel-out products.
  5. Overnight ligation of pACYC177 and OmpA_alpha.
  6. Overnight ligation of pACYC177 and OmpA_omega.

Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin

We have just received corrected version of omegaP-link10-homo2 primer. Let's hope this time everything will be fine. We are repeating the PCRs:


PCR
ProductTemplatePrimersProduct length
linker-ApDRIVE-TapTagAL+link10+homo2 and AP+NotI 470 bp
omega-linkerpUC19OmegaL+SacI and OmegaP+link10+homo2400 bp
TemperatureTime
94°C4:00
94°C0:3028 cycles
gradient 48-55°C0:45
72°C0:50
72°C10:00
4°Cinfinite