From Wiki | Meyer Lab
A protocol for quantifying dsDNA using a dsDNA specific dye binding assay that is unaffected by common contaminants e.g. RNA or oligonucleotides.
Step by step instructions
Still a work in progress
- Prepare DNA standards starting with a stock solution of calf thymus DNA at 250 ng/µl.
- Prepare working dye dilution.
- 1 µl dye, 1 µl enhancer, and 100 µl dilution buffer per sample or standard.
- keep dye cold and dark.
- Set up the plate.
- Add 100 µl diluted dye to each well
- then add 2 µl each sample or standard
- Measure fluorescence.
- Ex = 350, Em = 460
- Calculate concentrations.
- require that R^2 > 0.95
- and that your sample fluorescence is within the linear range of the curve
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Created 18:15 Apr 07, 2016 By: EliMeyer