Using the realtime PCR machine to measure GFP expression

As it turns out, the settings for reading the SYBR green dye also works for fluorescing GFP meaning that the realtime PCR machine in McM221 can be used in this way. Tried a quick test to see if I could get it to work. 4 samples monitored over the course of an hour at 37 deg:

• 30 ul LB broth uninoculated
• 30 ul LB + E.coli HB101 
• 30 ul LB + E.coli HB101 containing the pGLO plasmid, harvested off of an LB+Amp+Ara plate
• 30 ul LB + E.coli HB101 containing the pGLO plasmid, harvested off of an LB+Amp+Ara plate, but then also amended with 5 ul of 0.4 mg/ml arabinose. 
• The A580 reading for the E.coli without the plasmid was 0.29 which translates roughly into 1E8 cells/ml
• The A580 reading for the E.coli with the plasmid was 0.33 which translates roughly into 1.1E8 cells/ml
• Made a program to hold the temperature at 37 deg C and then to take fluorescence readings every minute for an hour.

Things to note from the graph

• bacteria without pGLO showed same level of fluorescence as just the media
• bacteria with pGLO were consistently higher
• bacteria without pGLO both showed  steady levels of signal across an hour while bacteria with pGLO increased. My assumption is that since the bacteria with the plasmid came from a plate that already had arabinose on it, the bacteria already were expressing GFP. Increases across time indicate the increase in GFP - could be due to cell doubling or due to increased GFP expression. Either way the pGLO cells without arabinose shouldn't have shown an increase. 

At the end of the day this shows promise as a tool for us to develop labs around, and of course research too.