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Measure femtogram quantities of dsDNA

Ben Ferneyhough, Nick Parkinson SBL UK 01/2011

In the modern molecular biology laboratory, DNA samples are a precious commodity, where only small amounts can be sacrificed for quality control analysis, preserving the majority for downstream applications such as Next Generation Sequencing. This requires the quantitation of femtogram DNA levels using low volumes (10 μL). With the Quant-iT Picogreen reagent Invitrogen provides a solution for measuring low concentrations of DNA. This, together with Cambridge Biosciences AccuBlue enhancer, makes it a highly sensitive fluorescence assay for dsDNA detection.

 

This application note investigated the use of very low volume DNA samples in a 384 well plate format on a BMG LABTECH filter-based microplate reader using the PicoGreen reagent in combination with Cambridge Biosciences AccuBlue enhancer. The sample volume was reduced to 10 µl while detection was possible down to 0.39 pg/µl.

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