Team:Tuebingen/Notebook/Protocols/yeast-trafo

From 2013.igem.org

(Difference between revisions)
Line 31: Line 31:
   <li>Resuspend cell pellet in 1000 µL YPD and plate 100 µL directly on appropriate selective plates.</li>
   <li>Resuspend cell pellet in 1000 µL YPD and plate 100 µL directly on appropriate selective plates.</li>
   <li>Colonies appear after 2 days of incubation at 30 °C.</li>
   <li>Colonies appear after 2 days of incubation at 30 °C.</li>
-
  <li></li>
 
</ol>
</ol>

Revision as of 03:25, 4 October 2013

Return to iGEM Main Page.

Yeast Transformation
Back to Protocols

 

Procedure

  1. Scrape some yeast cells off of a fresh YPD plate and inoculate in liquid YPD (= culture).
  2. Thaw ssDNA ("helper DNA).
  3. Pellet 1 mL of culture by centrifugation at > 13.000 rpm.
  4. Discard supernatant and resuspend cells in 100 µL ONE-STEP buffer. Vortex heavily.
  5. Add 20 µg ssDNA (10 µL of 2 mg/mL) and 100 - 500 ng plasmid DNA to be transformed. Vortext and incubate at 45 °C for 2 h.
  6. Add 1 mL YPD, mix and spin 10 sec at full speed. Discard supernatant.
  7. Resuspend cell pellet in 1000 µL YPD and plate 100 µL directly on appropriate selective plates.
  8. Colonies appear after 2 days of incubation at 30 °C.


Retrieved from "http://2013.igem.org/Team:Tuebingen/Notebook/Protocols/yeast-trafo"