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Nucleic Acid Quantification

dsDNA Assays


Helixyte™ Green Fluorimetric dsDNA Assay Kits (Cat# 17650 & 17651) simplify DNA quantification without sacrificing sensitivity. The assay provides a linear detection range between 0.2 ng and 1000 ng double-stranded DNA (dsDNA) (Figure 1.). The high-sensitivity DNA assay is ideal for quantifying PCR products, viral DNA, DNA fragments for subcloning and other applications requiring small amounts of DNA. Helixyte™ Green Fluorimetric dsDNA Assay Kits are highly selective for dsDNA over RNA and other common contaminants, including free nucleotides, salts, solvents and proteins. The Helixyte™ Green assay is an excellent replacement for PicoGreen®-based DNA assays.


The quantification of calf thymus DNA with Helixyte™ Green vs. PicoGreen®
Helixyte™ Green dsDNA Quantifying Reagent (Cat# 17597) can accurately quantify as little as 100 pg/mL of dsDNA using a fluorometer or 300 pg/mL using a fluorescence microplate reader. Helixyte™ Green dsDNA quantification assay is 10,000 times more sensitive than conventional UV absorbance measurements at 260 nm and at least 400 times more sensitive than the Hoechst 33258 dye based assay. Helixyte™ Green dsDNA Quantifying Reagent shows >1000-fold fluorescence enhancement upon binding to dsDNA, and much less fluorescence enhancement upon binding to single-stranded DNA (ssDNA) or RNA, making it possible to quantify dsDNA in the presence of equimolar amounts of ssDNA, RNA or proteins. Helixyte™ Green dsDNA Quantifying Reagent shows little if any AT- or GC-selectivity, enabling accurate DNA quantification.
 

RNA Assays


StrandBrite™ Green Fluorimetric RNA Quantitation Kit (Cat# 17655) provides a homogeneous assay for quantifying RNA in the presence of DNA. This RNA assay exhibits a linear detection range between 5 ng and 100 ng RNA (Figure 2.). Assay linearity is maintained even in the presence of several interfering compounds commonly found in nucleic acid preparations, including salts, urea, ethanol, chloroform, detergents, proteins and agarose. Its relatively high selectivity for RNA over dsDNA enables accurate RNA quantification in the presence of DNA and other common contaminants, including free nucleotides, salts, solvents and proteins, making this assay ideal for measuring samples for microarray, RT-PCR and northern blot procedures.


The quantification of RNA with StrandBrite™ Green Fluorimetric RNA Quantitation Kit (Cat# 17655).
StrandBrite™ Green Fluorimetric RNA Quantitation Kit allows detection of as little as 10 ng/mL RNA with a standard fluorometer, fluorescence microplate reader or filter-based fluorometer using standard fluorescein excitation and emission settings. The sensitivity is at least 20-fold better than that achieved with ethidium bromide and at least 100-fold better than that achieved using conventional absorbance measurements at 260 nm. Unlike UV absorbance measurements at 260 nm, StrandBrite™ Green RNA Quantifying Reagent (Cat# 17610) does not detect significant sample contamination caused by free nucleotides. Thus, StrandBrite™ Green RNA Quantifying Reagent more accurately measures the amount of intact RNA polymers in potentially degraded samples.
 

Table 1. Fluorescent Probes for Quantifying DNA & RNA Samples in Solution

Cat No.
Product Name
Ex (nm)
Em (nm)
Unit Size
Price
17597Helixyte™ Green dsDNA Quantifying Reagent *200X DMSO Solution*5015201 mL$327
17651Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *High Sensitivity*501520200 Tests$217
17650Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Microplate Readers*501520200 Tests$217
17655StrandBrite™ Green Fluorimetric RNA Quantitation Kit *Optimized for Microplate Readers*5005251000 Tests$327
17610StrandBrite™ Green RNA Quantifying Reagent *200X DMSO Solution*5005251 mL$327