Is there a way to separate free dye from fluorescently Labelled DNA?

01 January 2014 1 9K Report

I need to conduct a FRET analysis with labelled plasmid DNA and I am using YOYO-1 dye. I need to remove the free dye from the solution and get only the labelled DNA.

Is there any technique with which I can isolate or remove the free dye?

Martin Hengesbach

There are quite a number of ways to go:

An easy protocol would be precipitation of the DNA, using Ethanol or isopropanol. After two rounds, very little free dye should be present.

There are also commercially available spin columns, e.g. DyeEx by Qiagen; if you have a Plasmid, a Miniprep column purification will aslo work.

Last, small gravity flow size exclusion columns (NAP from GE) will also separate the dye from your DNA.

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