Composition 2X Loading dye for RNA electrophoresis
95% formamide
0.025% SDS
0.025% bromophenol blue
0.025% xylene cyanol FF
0.025% ethidium bromide
0.5 mM EDTA.
I wanna to prepare 10 ml of the solution
Am I right that I should add 4,9 ml pure water? And the more importantly How should I calculate the amount of EDTA that I should add? (I have 0.5 M EDTA stock solution made of EDTA disodium salt, dihydrate)
Calculate the volume using V1N1=V2N2 formula. N1 is the stock concentration which is 0.5M, V2 is the final volume (10ml), N2 is the final concentration (0.5mM). Calculate V1 which is how much volume you would take from stock to make the working concentration.
Add small quantity of water initially and add all the ingredients. After mixing all of them, make the final volume using water.
No, you do not start by adding 4.9 ml of water.
You need to calculate how much of each component you need to add, then add as much water as needed to make 10 ml.
Here's an easy example for your EDTA
V1 = V2C2/C1
V1 = (10 ml)(0.5 mM)/(0.5M) = 10 microliters
You should talk with a lab colleague to double-check your math before you mix anything, especially since ethidium bromide and formamide are both toxic.
Good luck!
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