4. You have 26 ul of RNA that you would like to load on a gel. You need to add a

Question

4. You have 26 ul of RNA that you would like to load on a gel. You need to add a gel loading buffer (which contains glycerol and a dye) so that the RNA sample sinks into the well. The gel loading buffer comes in a 5X concentrated stock. How much of the 5X gel loading buffer do you add to your sample? (keep in mind the volume of 5X loading buffer will increase the total volume - make sure you take this into account). 5. You wish to measure the concentration of a protein sample. When you measure the A2so, you get a reading of 1.92 from the spec. Since this is above the linear range of the spec, you take 20 1oftheprotein sample and add 180 l of water to it. The A280 of this new, diluted sample is 0.393. a) What is the concentration of protein in the diluted sample? b) What is the concentration of protein in the original, undiluted sample? c) If you wanted to load 10 ug of protein on a gel to analyze it, how many microliters of the original, undiluted protein sample would you use?

Explanation / Answer

10X V1= 1X 850/1000

v1= 0.85/10

v1= 0.085 L or 85 ml

therefore, 85 ml of 10 x stock is required make it upto 850 ml by adding 765 ml of D.H2O.

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