June 28
From 2007.igem.org
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Experiments
For [A]
- Seven dilutions of AI medium for pLLuxR.YpRC were made with 50 old:50 new 2xM9 along with -ve control: K12Z1 (1000 uM IPTG).
- One dilution (500 uM) of K12Z1 medium for pLLuxR.YpRC with 50 old:50 new 2xM9 was made.
- One dilution (500 uM) of new M9 medium for pLLuxR.YpRC was made.
For [B]
- The following inductions were done in 50 mL Glu M9 with [aTc]=10 ng/mL (10*50=0.5 ug == 50 uL of 10 ug/mL aTc)
pT LuxI.C: 2 flasks - 1 ul/mL == 50 uL
- 0.1 ul/mL == 5 uL
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- pL.LuxR.Y.pR.C and K12z1 were inoculated in LB.
- The required OD flask (0.205) was filtered.
- Seven dilutions for pL.Lux.R.YpR.C with 1:1 (AI supernatant:new 2xM9) were made in two sets along with -ve control: K12z1; AI+new M9 med;(500 uM IPTG) (2 tubes).
- Also 1 dilution of new M9 medium with pLLuxR.YpRC was made (two sets).
For [C,D]
- Inoculated pT.LuxI.C in LB.
Microcopy
- pT.LuxI.C that was growing in the flask before filtering was imaged.
- pL.cfp imaging was done.
- Open Loop samples [A] imaging was done.
