How to set up firefly bioluminescence experiment?
Posted: Tue Mar 19, 2019 9:34 am
Hey,
I was wondering if you can help me with setting up an experiment with regard to bioluminescence especially with the variables/factors involved in it.
I currently have several mg/ml of D-luciferin, luciferase (firefly), ATP solution. Furthermore I also got some MgSO4, but I don't know if I am going to need it or not. If I do need it and you think I can better replace it with another metal ions, please just tell me.
The experiment will be done (preferably several times) in a cuvette of 2,5 mL because I got to work with small amounts since the costs are pretty high.
The problem is that I don't know how I should begin, because I am struggling with a lot of factors like the ratio's of the chemicals. How much D-luciferin, luciferase, how much solvent (=water at the moment) do I need, how much ATP solution do I need to add.
I don't know whether I should dilute the solution or not.
Also, the optimal pH unknown as well and I dont know how to change pH without just random guessing how much of an acid/base I should add to get to the optimal pH.
At last IF the experiment works, the cuvette will be placed in a spectrophotometer with a range up till above a wavelength of somewhere above 1000 nm, but I have genuinely no idea what the proper way of calculating the ratio's, pH, whether I need to dilute it etc.
If you can guide me through it or give me tips about how I can deal with the problems, I would greatly appreciate it.
I was wondering if you can help me with setting up an experiment with regard to bioluminescence especially with the variables/factors involved in it.
I currently have several mg/ml of D-luciferin, luciferase (firefly), ATP solution. Furthermore I also got some MgSO4, but I don't know if I am going to need it or not. If I do need it and you think I can better replace it with another metal ions, please just tell me.
The experiment will be done (preferably several times) in a cuvette of 2,5 mL because I got to work with small amounts since the costs are pretty high.
The problem is that I don't know how I should begin, because I am struggling with a lot of factors like the ratio's of the chemicals. How much D-luciferin, luciferase, how much solvent (=water at the moment) do I need, how much ATP solution do I need to add.
I don't know whether I should dilute the solution or not.
Also, the optimal pH unknown as well and I dont know how to change pH without just random guessing how much of an acid/base I should add to get to the optimal pH.
At last IF the experiment works, the cuvette will be placed in a spectrophotometer with a range up till above a wavelength of somewhere above 1000 nm, but I have genuinely no idea what the proper way of calculating the ratio's, pH, whether I need to dilute it etc.
If you can guide me through it or give me tips about how I can deal with the problems, I would greatly appreciate it.