IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/05/26: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 26: Line 26:
* Tetracycline and Kanamycin construction plasmids - 1 µL of DNA into competent cells stock
* Tetracycline and Kanamycin construction plasmids - 1 µL of DNA into competent cells stock
* On ice for 30 minutes
* On ice for 30 minutes
* Heat shock at 42ºC for 45 seconds
* Heat shock at 42.5ºC for 45 seconds
* 2 minutes on ice
* Added 400 µL of sterile LB broth to the 100 µL of glycerol stocks (in the same tube as the glycerol stocks)
* Recovery for 2 hours at 37ºC


<!-- ## Do not edit below this line unless you know what you are doing. ## -->
<!-- ## Do not edit below this line unless you know what you are doing. ## -->

Revision as of 14:18, 26 May 2009

iGEM Project name 1 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Testing Chloramphenicol Plates

  • Cells grew in both LB-only and LB+Chlor
  • Possibly due to insolubility of antibiotic.
  • Made LB Broth only (10 mL) and LB Broth + Chloramphenicol (10 mL Broth + 10 µL Chloramphenicol)
  • Innoculated with DB3.1 cells
  • Left to grow in Lagally Lab shaker till next morning.

Obtained results on Tetracycline plate

  • retrieved plates at 13:06 from the incubator from Lagally Lab BioHazard room
  • observed no growth on LE-tet plate
  • observed cell growth on LE-only plate
  • Conclusion: Tetracycline is effective

Transformation of Biobricks

  • The GFP, pBAD, Terminator and J23100/RBS showed slow growth colonies, only becoming apparent this morning.
  • The Tet Construction Plasmid and Kan Construction Plasmid showed no growth, will perform another transformation of these Biobricks

Transformation of Tet and Kan Construction Plasmids

  • Transformation performed in DB3.1 cells
  • Tetracycline and Kanamycin construction plasmids - 1 µL of DNA into competent cells stock
  • On ice for 30 minutes
  • Heat shock at 42.5ºC for 45 seconds
  • 2 minutes on ice
  • Added 400 µL of sterile LB broth to the 100 µL of glycerol stocks (in the same tube as the glycerol stocks)
  • Recovery for 2 hours at 37ºC


Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:British_Columbia/2009/Notebook/Biosensor_Sensitivity/2009/05/26&oldid=310356"