IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/06/13

From OpenWetWare
Jump to: navigation, search
Igem-logo-150px.png 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>      </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Preparation of pUC 19 (Done with Hank)

  • Took out ON culture of pUC19-containing cells (innoculated on 12 June 2009).
    • Note that the culture rotor was not spinning - tube may not have been well-aerated.
  • Mixed the entire 80µL of RNaseA into Charge-Switch Pro Resuspension buffer.
  • Aliquoted 1.25 mL of ON culture each per microfuge tube (4 in total), spun down at max speed for 10 min.
  • Followed ChargeSwitch Pro miniprep protocol from here on; combined the harvested cells into two tubes (one from Hank, one from me).
  • Nanodropped the samples:
Tube [DNA] (ng/µL) A260/280
pUC19 (1) 31.11 1.26
pUC19 (2) 26.36 1.27
  • Stored at AMBLE -20ºC freezer
  • Moved to Lagally Lab -20ºC freezer, labelled:
    • pUC19 HYEM(1)
    • pUC19 HYEM(2)
  • Made glycerol stocks of these cells (250µL of 60% sterile glycerol + 750µL of ON culture)

Results of Mutagenesis

  • Hank noticed that the media was cracked; could be too thin.
  • Took plates out from AMBL 37ºC incubator @ 2:45 PM.
  • Observations
Plate Colony count Colors
Positive (conc.) 0 n/a
Positive (dil.) 0 n/a
Negative (conc.) TNTC White (all)
Negative (dil.) 135 White (all)
Control (dil.) 20+1 Blue+White
Control (conc.) 322+33 Blue+White
  • Plates were parafilmed and stored at AMBL4ºC.