Flourometer and Spectrophometer, which give accurate result? - (May/14/2007 )
Any has experiences on using spectrophometer and flourometer for measuring DNA and which one give better accurate results?
Thanks in advance
Both are commonly used. Specy is fine for DNA purity work (though in my lab we didn't always bother) using the 260/280nm ratio. I think you'd need fluoresence if you wanted to sequence the DNA, though. Fluoresence, with current instrumentation, has the sensitivity which specy doesn't given the low DNA concentrations found. It might not be long before we can sequence on specy, though.
Thanks paraboxa, but my question is just know which instrument could give accurate result on DNA conventration.
Specy is adequate using the 260/280 ratio.
Agreed.... I also use spectro and it works fine so far.
Fluorometer is far more accurate, but you need the expensive dye that goes with it and you won't see if there is any contamination (because of the dye's specificity),
whereas spectro is enough as a routine and also gives potential contaminations indication.
To give you an idea: for calibrating DNA samples before quantitative PCR, I would choose fluo, otherwise spec is more than enough.
But mostly, in the end you don't care so much, because even if you get the exact concentration of your DNA or RNA or whatsoever, with measurements reproduced 10 times, your hand will never be accurate enough to pipet the right volume when you will need to.
For your purpose spec. is enough and round up the decimals to only one.