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After adding the volumes of 2X master mix to their sample tube and control tube each,...

After adding the volumes of 2X master mix to their sample tube and control tube each, the student decides to add 5ul of DNA + water to dilute the reaction to it's final 1X concentration. How much water should the student add to do so?

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Answer #1

Standard PCR reaction can be set from volume 25uL, 30uL, 40uL, 50uL upto 200uL per tube. Any reaction volume above 200uL needs to be either split into small reactions or separate 0.5ml tube should be used.

Considering on an avg if the student is going to set up 40uL of PCR reaction, reagent volume will be as below:

Note: using the below values one can always calculate by cross multiplication how much water should be added based on their reaction volume

Reaction voume 20uL 40uL 100uL

2X Master mix - 10uL 20uL 50uL

Vol of DNA -5uL 5uL 5uL

Vol of water - 5uL 15uL 45uL

Total volume- -20uL 40uL 100uL

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