What is Eva Green dye?

What is Eva Green dye?

Product Description. EvaGreen® Dye is a patented green fluorescent nucleic acid dye that is essentially non-fluorescent by itself, but becomes highly fluorescent upon binding to dsDNA.

Why does SYBR Green dye not work well in high resolution melting HRM analysis?

Intercalating dyes SYBR Green I is a first generation dye for HRM. It fluoresces when intercalated into dsDNA and not ssDNA. This is because it is suggested that the lack of accuracy may result from “dye jumping”, where dye from a melted duplex may get reincorporated into regions of dsDNA which had not yet melted.

What is HRM qPCR?

High resolution melt analysis (HRM analysis) is the quantitative analysis of the melt curves of product DNA fragments following PCR amplification. HRM can be considered the next-generation application of amplicon melting analysis. Related Topics: qPCR Assay Design and Optimization and qPCR/Real-Time PCR.

Is EvaGreen an intercalating dye?

EvaGreen is a new DNA intercalating dye successfully used in quantitative real-time PCR. Good separations of dsDNA fragments were obtained using the three dyes.

Does SYBR Green bind to ssDNA?

All SYBR dyes bind to dsDNA, ssDNA and RNA but vary in sensitivity. SYBR Safe DNA Gel Stain was specifically developed as a safer alternative to ethidium bromide. SYBR Green I is an ultrasensitive stain for dsDNA, and SYBR Green II is a highly sensitive stain for RNA and ssDNA.

How does SYBR Green work in qPCR?

SYBR Green is one of the most commonly used fluorescent dyes in qPCR. It binds to double-stranded DNA molecules by intercalating between the DNA bases. Once intercalated to DNA, SYBR Green becomes less mobile, causing its energy to be released as fluorescence.

What does melt curve tell you?

Melting curve analysis is an assessment of the dissociation characteristics of double-stranded DNA during heating. The temperature at which 50% of DNA is denatured is known as the melting temperature. The information gathered can be used to infer the presence and identity of single-nucleotide polymorphisms (SNP).

What is an intercalating dye?

Intercalation occurs when ligands of an appropriate size and chemical nature fit themselves in between base pairs of DNA. These ligands are mostly polycyclic, aromatic, and planar, and therefore often make good nucleic acid stains.

What is the difference between SYBR Green and Taqman?

Key Difference – SYBR Green vs Taqman SYBR Green is a method based on intercalating nucleic acid staining dye while Taqman is a method based on hydrolysis probe. Both technologies are designed to generate fluorescence during the PCR, which allows real-time PCR machine to monitor the reaction in “real time”.

What are the major differences between the Taqman and SYBR Green method of qPCR?

Key Difference – SYBR Green vs Taqman SYBR Green and Taqman are two methods employed to detect or watch amplification process of real-time PCR. SYBR Green is a method based on intercalating nucleic acid staining dye while Taqman is a method based on hydrolysis probe.

Are EvaGreen® dyes suitable for real-time PCR applications?

The unique properties of Biotium’s EvaGreen® dyes make them particularly useful in quantitative real-time PCR applications. Relative to other green dyes, such as SYBR® Green I, both EvaGreen® dyes are generally less inhibitory toward PCR and less likely to cause nonspecific amplification.

What is the highest concentration of EvaGreen dye?

Highly concentrated EvaGreen® Dye, 2000X (2.5 mM) in DMSO is offered for customers with specialized applications requiring a concentrated dye solution. For most qPCR applications, we recommend our ready-to-use Forget-Me-Not™ EvaGreen® qPCR Master Mixes or EvaGreen® Dye, 20X in water.

Is EvaGreen® dye superior to SYBR® Green I?

The result of our efforts is a dye superior to SYBR® Green I and other commercial PCR or high-resolution melt curve (HRM) dyes.* EvaGreen® Dye is used in all of Biotium’s dye-based qPCR master mixes, which combine superior brightness and sensitivity, with the ability to do sensitive melt curve analysis in the same reaction.

What is EvaGreen used for in qPCR?

In addition to qPCR and HRM, EvaGreen® Dye is currently the only qPCR dye to be used in droplet digital PCR (ddPCR). EvaGreen® can also be used for isothermal amplification, microfluidic PCR systems, capillary gel electrophoresis, and more.