Promega Corporation

Quantitating DNA from FFPE tissue: Superior sensitivity of the QuantiFluor™ dsDNA...

Quantitating DNA from FFPE tissue: Superior sensitivity of the QuantiFluor™ dsDNA System versus the NanoDrop Spectrophotometer Poster

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Abstract

Accurate quantitation of DNA concentration is critical for many applications. Traditional spectrophotometric assays cannot determine DNA concentrations below 2µg/ml; however, many isolated DNA samples have concentrations well below this level. The QuantiFluor™ dsDNA System (Cat# E2670) is a fast, easy and sensitive method for determining low DNA concentrations.

Formalin fixation and paraffin embedding is the most commonly used method worldwide for tissue storage. The Maxwell®16 FFPE Tissue LEV DNA Purification Kit (Cat# AS1130) is specifically designed for optimal purification of DNA from one to ten (5µM) sections of FFPE tissue samples. To help minimize PCR inhibitor carryover, small amounts of the DNA-binding paramagnetic particles are used, and as a result the DNA-binding capacity of the system is limited to a few hundred nanograms of pure DNA suitable for amplification applications including qPCR. With such low levels of DNA, an accurate and sensitive method is required for DNA concentration determination.

Double-stranded DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples using the Maxwell® 16 FFPE Tissue LEV DNA Purification Kit (Cat# AS1130) was quantitated using both the Quantifluor™ dsDNA System and NanoDrop spectrophotometer. The NanoDrop was only able to determine concentrations for samples above 2ng/µl but Quantifluor™ accurately determined concentrations for all samples.

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  • Part# PS122
  • Printed in USA 3/11.

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