Picogreen Microtiter Plate Assay

As higher throughput methods for DNA determination are being developed, more automated procedures are needed for rapid assessment of extracted DNA quantity prior to DNA amplification. To this end, the Forensic Science Service has developed a PicoGreen assay that is capable of detecting as little as 0.25ng/mL of double-stranded DNA in a 96-well microtiter plate format (Hopwood et al. 1997). PicoGreen is a fluorescent interchelating dye whose fluorescence is greatly enhanced when bound to double-stranded DNA.

To perform this microtiter plate assay, 5 | L of sample are added to 195 | L of a solution containing the PicoGreen dye. Each sample is placed into an individual well on a 96-well plate and then examined with a fluorometer. A 96-well plate containing 80 individual samples and 16 calibration samples can be analyzed in under 30 minutes (Hopwood et al. 1997). The DNA samples are quantified through comparison to a standard curve. This assay has been demonstrated to be useful for the adjustment of input DNA into the amplification reaction of STR multiplexes (Hopwood et al. 1997). It has been automated on a robotic workstation as well. Unfortunately, this assay quantifies total DNA in a sample and is not specific for human DNA.

Stuttering Simple Techniques to Help Control Your Stutter

Stuttering Simple Techniques to Help Control Your Stutter

Discover Simple Techniques to Help Control Your Stutter. Stuttering is annoying and embarrassing. If you or a member of your family stutters, you already know the impact it can have on your everyday life. Stuttering interferes with communication, and can make social situations very difficult. It can even be harmful to your school or business life.

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