I carried out a DNA extraction with purification using magnetic beads. The eluate had a lot of bead residue.
We removed the beads, placed the samples on an agarose gel and observed genomic DNA bands. But when quantifying the samples by Fluorescence, the quantification was zero/negative?
Which reagent can interfere with DNA quantification by fluorescence?
Has anyone experienced this?
What type of dye are you using? you can also try other quantification methods
Can you tell me what buffer or solution your DNA is dissolved in, and which quantification method and dye you used?
Possibilites due to: differences in dye specificity, buffer conditions, DNA integrity, and the presence of contaminants.
What do you mean by "quantifying by fluorescence"? Unless you've added labeled nucleotides or a dye like ethidium bromide, DNA is only very weakly fluorescent. Do you mean use measured concentration in a spectrophotometer?
The most likely issue is not zeroing the machine with the correct buffer.
Hi guys. Thank you for your attention.
We used Qubit to quantify the samples and they were eluted in the kit's elution buffer, which we believe is TE Buffer.
We believe that the residue of the beads influenced the fluorescence.
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