Biotium expands multiplexing options for probe-based qPCR with unique red qPCR dye

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EvaRuby™ Dye offers minimal cross-talk in neighboring channels and is suitable for probe-based qPCR with HRM® analysis in the same reaction.

Caption: High Resolution Melt Data Difference Plot. EvaRuby™ Dye at 1X in Forget-Me-Not™ Universal Probe qPCR Master Mix (Cat. No. 31043) was used to detect the Applied Biosystems® MeltDoctor™ HRM Positive Control Kit. Data was acquired using the Qiagen® Rotor-Gene® Q, 0.1°C/2 sec, with a 470 nm excitation and 610 nm emission filter. Data analysis was performed with uAnalyzeSM (https://dna-utah.org) using baseline subtraction. EvaRuby™ Dye can clearly distinguish each homozygous variant as well as the heterozygous variant.

Fremont, CA – Biotium, a leading life science reagent supplier and inventor of the widely used EvaGreen® qPCR Dye, announces the release of EvaRuby™ Dye for qPCR and HRM®. This spectrally unique red fluorescent intercalating dye can be incorporated into probe-based qPCR assays, expanding multiplexing options for probe-based qPCR. EvaRuby™ Dye is compatible with ~470 nm excitation and ~610 nm emission filters.

While a variety of DNA binding dyes with different fluorescence emission properties have been published for use in qPCR and HRM®, only the green fluorescent dyes SYBR® Green I, SYTO™ 9, and EvaGreen® Dye have widespread use in these applications. EvaGreen® Dye is currently the only dye suitable for qPCR and HRM® in the same reaction, however, due to its relatively broad emission, EvaGreen® cannot be combined with red PCR probes for multiplexing or HRM® in probe qPCR reactions. As a solution, Biotium scientists have developed EvaRuby™, a unique dye with a much longer Stokes shift of 480/613 nm (excitation/emission). This allows excitation of EvaRuby™ Dye by the same green/blue channel as EvaGreen® Dye, and detection in the orange (JUN™ or ROX™) channel. As a result, EvaRuby™ Dye has minimal cross-talk in standard probe detection channels and enables the combination of dye-based and probe-based qPCR in a single reaction. In addition, EvaRuby™ Dye is validated for HRM® analysis and may be used as an internal control for probe-based qPCR to detect non-specific amplification that goes undetected by target-specific probes. EvaRuby™ Dye is also suitable for singleplex qPCR, LAMP, and gel electrophoresis.

EvaRuby™ Dye has been validated on Applied Biosystems® QuantStudio™ 5 96-well and Qiagen® Rotor-Gene® Q instruments, with potential compatibility for other qPCR instruments capable of acquiring data using ~470 nm excitation and ~610 nm emission filters. EvaRuby™ Dye¬ is available as a 20X in water solution in standard 2×1 mL and 500 uL trial sizes priced at $320 and $90 respectively.

Features and benefits of EvaRuby™ Dye include:

  • Spectrally Unique: Minimal cross-talk in neighboring channels
  • Flexible: Unique red dye that expands multiplexing capabilities for probe-based qPCR
  • Versatile: Can combine probe-based qPCR with HRM® in the same reaction
  • Convenient: Easily confirm your results with melt curve analysis rather than time consuming post-qPCR gel electrophoresis

The release of EvaRuby™ Dye adds to Biotium’s industry-leading catalog of qPCR reagents for life science research. For more information on EvaRuby™ Dye, please visit the product page. For licensing inquiries, please contact btinfo@biotium.com.

About Biotium

Biotium is a leading life science reagent manufacturer and supplier devoted to providing high-quality and innovative fluorescent tools that fuel scientific discovery. Our collaborative team of experienced chemists and biologists, who are at the forefront of fluorescent dye design, apply chemistry-based principles toward producing solutions for unmet challenges in life science and medical research. Since its founding in 2001, Biotium has developed over 30 patented technologies that have been licensed out to leading life science technology companies worldwide. For more information, please visit www.biotium.com.
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