Loading Dye in Q5 PCR product?

15 July 2021 1 7K Report

Hi everyone,

I have a question that might sound silly. I'm performing a PCR with Q5 polymerase, in a reaction volume of 25mL. Now, I would need to run all of the volume of the reaction to excise a band. The question is, how much loading dye (I'm using a 6X TriTrack) should I add to visualize the bands in the gel?

Thanks!

Pierre Béguin

As mentioned in the instructions provided by the supplier, add 1 volume of 6X loading buffer for 5 vol. of sample. In your case, 5 ul for 25 ul of sample. Just make sure that the well in the gel is large enough to accommodate 30 ul, if need be by taping together two or more teeth of the well-forming comb.

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