DNA Quantitation
The detection and quantitation of small amounts of DNA is extremely important in a wide variety of applications. The most commonly used technique for measuring DNA concentration is determining the absorbance at 260 nm. However, this method has major disadvantages, including:
- the relative contribution of nucleotides and single-stranded nucleic acids to the signal.
- the interference caused by contaminants found in nucleic acid preparations.
- the inability to distinguish between RNA and DNA.
- the relative insensitivity of the assay.
The Core Facility offers 2 highly sensitive and specific forms of fluorescent based methods for the detection of dsDNA:
- PicoGreen (Invitrogen/Molecular Probes)
- Quantifiler (Applied Biosystems)