Melbourne/Lab Notebook gv F1


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Testing Floatation

  • Growing up cells from glycerol stocks overnight in 5mL LB amp
  • Prepare 250ml conical shaker flasks by cleaning and autoclave.
  • Add 50mL of LB Amp to each of two flasks.
  • Add 1M IPTG to one flask to make final concentration of 1mM IPTG, Lable with I.
  • Transfer 1mL of overnight culture to each flask.
  • Grow overnight at 37degC on a shaker.
  • Next morning transfer to 50ml falcon tubes.
  • Spin down the cells gently 25minutes at 1000rpm in Eppendorph 5810R centrifuge.
  • Poor off and discard the LB, maintaining the pellet and a residual ammount of LB 1ml about.
  • Replace LB with 50ml of 10g/L NaCl filtered through a 0.22um filter.
  • Resuspend the cells by shaking.
  • Leave at room temperature and observe the settling over the next week.

Example pNL29(orininal plasmid) and Tube(64) the IPTG promoted version of BBa_I750016

Image:Melbourne-float1 11am 18-10-2007.JPG