Team:Warsaw/Calendar-Main/14 May 2008

From 2008.igem.org

(Difference between revisions)
Line 48: Line 48:
-
Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> (negative control) genomic DNA<br>
+
Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (negative control) genomic DNA<br>

Revision as of 19:10, 4 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
Interactive list of topics
next notebook entry

 


Michał K.:

  1. Setup of 9 separate cultures from 9 colonies of transformants pMPMT5+AID (liquid LB+tetracycline).
  2. Optimization of conditions for PCR to obtain AID DNA fragment - annealing temperature gradient (from 62°C to 82°C).
    Primers: AIDlNrH AIDpLinB
    Template DNA: pTrc99A-AID
    Elongation time: 60 sec
    20 cycles
  3. Optimization of conditions for PCR - T7 RNA polymerase for translation fusion; annealing temperature gradient (from 62°C to 82°C).
    Primers: T7lLinkB T7pXbSal
    Template DNA: E. coli Rosetta genomic DNA
    Elongation time: 4 minutes
    20 cycles
  4. Optimization of conditions for PCR - T7 RNA polymerase for transcription fusion; annealing temperature gradient (from 62°C to 82°C).
    Primers: T7lRBSHi T7pXbSal
    Template DNA: E. coli Rosetta and TOP10 (negative control) genomic DNA
    Elongation time: 4 minutes
    20 cycles
  5. Gel electrophoresis of PCR products.

Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/14_May_2008"