TIPS & TECHNIQUES Small Amount of dsDNA to Quantitate? You can do it with the Quantus™ and QuantiFluor® dsDNA System A ccurate quantitation of DNA is critical for many applications. Traditional spectrophotometric assays cannot determine DNA concentrations below 2ng/μl, and many DNA isolates have concentrations well below that level. Using the Quantus™ Fluorometer with QuantiFluor® dsDNA System provides a fast, easy and sensitive method for determining DNA concentration to as low as 10pg/μl from just 1μl of sample. The QuantiFluor® dsDNA System contains a fluorescent DNA-binding dye that enables sensitive and specific quantitation of small amounts of double-stranded DNA (dsDNA) in solution. The dye shows minimal binding to singlestranded DNA (ssDNA) and RNA. Using the QuantiFluor® dsDNA System, we have detected sample dsDNA concentrations as low as 10pg/μl using 1μl of sample input per assay. It is possible to quantitate more dilute samples by adding more sample per assay. Up to 100μl of sample may be measured per 200μl assay. This article describes a protocol for use of QuantiFluor® dsDNA dye with the Quantus™ Fluorometer. The Quantus™ Fluorometer measures sample volumes as little as 1μl in a 200μl assay volume without sacrificing instrument sensitivity. Measuring dsDNA concentration using the QuantiFluor® and the Quantus™ Fluorometer. 250 dsDNA System Fluorometer 200 150 y = 1.008x + 0.7839 r2 = 0.999 100 The standard curve demonstrates assay linearity. The values presented correspond to assay tube dsDNA concentration and not starting sample concentration. 50 0 050 100 150 Actual Concentration (ng/µl) 200 250 enews | July 2015 Expected Concentration (ng/µl) 11792MA Pagina 13
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