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

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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:
 * 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
 * Plates were parafilmed and stored at AMBL4ºC.


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