You may need a gfp standard curve to know for sure. Anything else would be back of the envelope. Would it be feasible to buy an isolate of your fluorophore or hisTag it and purify? Thete should be papers on measuring this but IMHO taking optical density readings of bacteria in stationary phase against a fluorescent protein standard curve using YOUR spectrophotometer (not all created equal) would be the most quantifiable.
Sebastian S. Cocioba
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
--Sure, I dont have it with me right know, but it was something like this:I inoculated colonies in LB and let them grow overnight. In the morning I diluted it in M9 medium and let it grow for 4 hours, then I did the first measurement (T0), I measured the OD and fluorescence. After another 1 hour of incubation I measured the OD and fluorescence again (T1).At T0 the fluorescence was around 50 and at T1 it was around 70, the OD I cant remember. It would be great to get an estimate of the GFP concentration inside the cell.Thanks!On Thursday, May 14, 2015 at 8:02:45 PM UTC-3, Linden wrote:Hi guys!I would like to rank in strength a few promoters I have. The promoters are cloned upstream GFP, so I used a spectrophotometer (485 nm excitation 525 nm emission) to measure the GFP in E. coli, just like the paper "Measuring the activity of BioBrick promoters using an in vivo reference standard". I did everything, and now I have a some numbers, but I don't know how to turn it in something useful. Anyone know which formula should I use to get a estimate of the GFP or mRNA concentration inside the cell?Thanks a lot!
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