Spectrophotometer is quick and easy to use but has a higher limit of detection and quantification than a fluorometer.
Dear Jochen, thank you very much for your reply. I tried a micro-spectrophotometer but I have had some problems to have good reproducibility of some plant samples vs. with a fluorometer (i.e. Qubit).
Now I would like to buy one but I must decide.
All the best, Olga
Yes, this I forgot: the fluorimeter is less suceptible to components that absorb around 260nm. Especially if you have contaminations with phenol or proteins a fluorescence measurement allows precise quantification whereas the spectrometer gives you a biased result.
Just for curiosity: is it the very same sample giving you different readouts on the spectrometer? This should actually not be the case... then the instrument is somehow "bad" or your handling is "suboptimal". If different samples give different readings, then it might be that you have differing amounts of contaminations (phenol, proteins) in these samples. Maybe a diffetent purification method can help.
Dear Joachen,
surely the strument was not good. Often the plant material has a lot of problems and when I use good kit I have not problem...but when I use a normal CTAB method It is possible to have not pure DNA (Polyphenols, mucilages and so on).
Thanks again. Olga
That's it. So if you suffer from such "bad material" and you cant improve the isolation method, then the fluorimeter is the better choice. However, be aware of the possibility that these contaminants may influence downstream reactions.
Spectrophotometer is the method of choice for detection of DNA and RNA but with flourimeter your material should be absolutely pure or else you get erroneous readings.
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