DNA quantity (Concentration) is very important step in pre-PCR technique.
Nanodrop is great for routine PCR work but I would suggest that you use fluorometric quantification (Quantus) for sensitive applications such as high-throughput sequencing. The dye used in fluorometers selectively bind to dsDNA and it can give a more accurate quantification, especially in lower concentrations (if I remember correctly,
Hello! I use Quantus to measure low concentrations of RNA or DNA (0.05 ng/microL and more). In compare to Quantus, Nanodrop doesn't require additional kits.I use Nanodrop for total RNA/DNA or plasmid measurments.
Nanodrop is easy to use and more accurate. Also requires just 1microlitre or so which is convenient in term of saving more of your samples. So my preference is nanodrop
Nanodrop is great for routine PCR work but I would suggest that you use fluorometric quantification (Quantus) for sensitive applications such as high-throughput sequencing. The dye used in fluorometers selectively bind to dsDNA and it can give a more accurate quantification, especially in lower concentrations (if I remember correctly,
Quantus is more sensitive because of a reagent specific, enabling I to quantify small concentrations through a kit with fluorochrome, since it is nanodrop is less sensitive when compared to Quantus, yet it tells you if your sample has contaminants like: Guanidine, Ethanol, Among others.
It is important that you quantify and qualify as samples of the two equipments.
I add either EtBr or Pico Green to a sample along with a DNA of known concentration creating a standard curve. These can be spotted directly on a UV Box for a ball-park number or if you have a fluorescent plate reader, you can generate hard numbers. Thermo Fisher sell a kit now called Quant-iT that sounds like it does the same thing. Pico Green should get you into the picogram range for sensitivity. Quant iT claims down to 25 pg sensitivity.
It depends on the purpose you will use your sample, for routine procediments: Nanodrop
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Comparação na quantificação de amostras DNA e RNA por meio de espectrofotometria e fluorimetria Tiago César Moreira · Luciana de Andrade AgostinhoArquivo disponível · Poster · Maio de 2017- 10.13140/RG.2.2.24760.44801
It depends on what you need it for. Routine quantification of DNA can be done with a Nanodrop,it is easy and as most already said do not require additional reagents and preparation. However, it is not as sensitive as it also picks up background (if your sample is dirty, protein and RNA) during the measurement where other methods rely on specific dyes that binds specifically to the DNA and the dye is measure instead of your DNA which makes it much more sensitive. For everyday DNA quantification a Nanodrop is fine.
Nanodrop is good for routine quantification or low volumen samples.
Quantus is good for small concentrations of DNA.
I used Quantus to quantify bacterial DNA for sequencing.
Quantus. Unless your DNA is column purified, you will get wildly inaccurate readings from nanodrop due to protein and/or polysaccharide contamination. Quantus and other fluorimetric measurement methods are not sensitive to such contaminants.
For doing normal PCR nanodrop is ok.
But for sequencing work and any other purpose i will suggest Quantus.
If you are interested in precise determination of the amount of genomic material available use the Quantus assay! Mark Farman gave a very good explanation why this should be the method of choice before. Spectophotometric readings give you additional information about the purity level of your extract(s) but the accuracy of determining NA concentration is highly influenced by possible contaminants using this technique. Good luck!
It is completely incorrect to say that Nanodrop is more accurate. These are both physicochemical methods for measuring biomolecules. If you have completely pure DNA, they will both be equally accurate unless your machine is out of calibration, or reagents/buffers/standards have gone bad. In fact, unless you can be absolutely certain that you do not have polysaccharide/protein or RNA/DNA contamination (depending on whether you're measuring DNA/RNA), fluorometry will always be more accurate because the intercalating fluorescent dyes will not bind to contaminants. On teh other hand, these molecules will absorb light in the 260/280 nm wavelengths. As a cautionary tale, even column purification will not get rid of certain polysaccharides from certain DNA preparations. Nanodrop is badly affected by polysaccharide contaminants, fluorometric methods are not.
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