Measurement of Nucleic Acid Concentrations Using the DyNA Quant™ and the GeneQuant™

Abstract

Molecular biology is now a routine tool in almost all biological research fields. With the exponential growth in the number of molecular biological techniques, there is a recognizable need for sensitive, accurate and precise quantitation of nucleic acids. We present here two complementary instruments designed for the quantitation of nucleic acids, the GeneQuant II and the DyNA Quant 200 Fluorometer. The GeneQuant II can rapidly determine the UV absorbance of a solution and display the calculated DNA, RNA or protein concentration. In addition, the GeneQuant can display calculated melting temperatures for a given DNA oligonucleotide base sequence, a useful feature for primer design. The DyNA Quant 200 quantitates DNA on the basis of the fluorescent Hoechst 33258 dye/double-stranded (ds)DNA assay. Upon binding to dsDNA, the spectral properties of the dye change such that it becomes highly fluorescent at 460 nm when excited at 365 nm. The assay has proven to be a specific and sensitive alternative method for DNA quantitation, particularly for unpurified DNA samples. Together, the GeneQuant II and the DyNA Quant 200 are a cost-effective and convenient solution to the routine protein and nucleic acid quantification needs of the molecular biologist.

Optimised cetyltrimethylammonium bromide (CTAB) DNA extraction method of plant leaf with high polysaccharide and polyphenolic compounds for downstream reliable molecular analyses

“…This leads to false overestimate of DNA concentration (Sambrook et al, 1989). However, it is still the simplest and fastest method of determining DNA concentration and purity (Teare et al, 1997). However, from the results, it can be seen that the modification steps were helpful in obtaining DNA of reasonable good yield and quality.…”

Microplate fluorescence assay for the quantification of double stranded DNA using SYBR Green I dye

“…The dsDNA binding dye Hoechst 33258 used in conjunction with the Hoefer DyNA Quant 200 Fluorometer can detect as little as 20 ng DNA in a 2 ml sample (Teare et al 1997). However, this dye has a preference for AT-rich sequences and is therefore sensitive to the DNA composition (Cesarone et al 1979;Gallagher 2004).…”

“…However, this dye has a preference for AT-rich sequences and is therefore sensitive to the DNA composition (Cesarone et al 1979;Gallagher 2004). Furthermore, the detectability of this method is limited by the length of the DNA sequence, as DNA fragments shorter than 200 bp may not have the minimum number of consecutive AT base pairs required for efficient Hoechst dye binding (Teare et al 1997).…”

Selección de marcadores microsatélites (SSR’s) para el análisis de variabilidad genética en siete cultivares de arracacha (Arracacia xanthorrhiza)

“…El análisis cualitativo del ADN extraído permitió obtener una banda definida de alta calidad, con algunas trazas de ARN o DNA degradado (figura 1), con concentraciones entre 136 a 565 ng/ ml y una relación A260/A280 calculada entre 1.78 a 1.93 estableciéndose que todas las extracciones presentaban una calidad óptima (Teare et al, 1997). Se usó en la mezcla de reacción una concentración de ADN de 20 ng/µL, concentración descrita por Morillo et al (2004).…”