How does SYBR Green 1 work?
SYBR Green I is a dsDNA binding dye, which can be used to quantify amplicon amount during the course of the PCR by tracking overall fluorescence emission. The dye binds into the minor groove of dsDNA, and does not bind to ssDNA. When bound, it increases its fluorescence by up to 100 fold (Figure 6).
What polymerase is in SYBR Green?
|For Use With (Equipment):||7000 System, 7300 System, 7500 System, 7700 System, 7900HT System,Applied Biosystems StepOnePlus™ Fast Real-Time PCR System, QuantStudio™, StepOne™, Standard Mode, StepOnePlus™, Standard Mode, ViiA™ 7 System|
|Polymerase:||AmpliTaq Gold DNA Polymerase|
What is master mix used for?
The master mix enables researchers to set up controls and test different concentrations of their target DNA or RNA templates without having to individually add precise amounts of enzymes, buffers, cofactor (usually MgCl2), water and dNTP to each reaction tube or plate well.
What is the role of SYBR Green?
SYBR® Green is a dsDNA-binding dye that intercalates nonspecifically into dsDNA, allowing measurement of the amount of PCR product. Since the increase in fluorescence is proportional to the amount of product accumulated, SYBR® Green qPCR can be used for relative DNA quantification.
What is qPCR Master Mix?
Master mixes for qPCR contain the necessary reagents for a straightforward quantitative PCR or real-time PCR reaction. A given mix will typically include the polymerase, buffer, dNTPs, MgCl2, and fluorescent dye. In concentrations from 2X to 5X, it is ready-to-use with the addition of template and primers.
Is SYBR Green toxic?
Mutagenicity of these stains was not observed although their toxic concentration was reached. Thus, SYBR Gold and SYBR Green II do not show mutagenicity in our tests, even at toxic doses, and these DNA stains represent safer alternatives to ethidium bromide for nucleic acid visualization.
What does SYBR Green do in qPCR?
SYBR® Green for qPCR Sso7d is a double-stranded (ds) DNA–binding protein that increases speed and processivity and increases tolerance to PCR inhibitors. SYBR® Green is a dsDNA-binding dye that intercalates nonspecifically into dsDNA, allowing measurement of the amount of PCR product.
What are the components of a master mix?
A PCR master mix is a premixed concentrated solution that has all of the components for a real-time PCR reaction that are not sample-specific. A master mix usually contains a thermostable DNA polymerase, dNTPs, MgCl 2, and proprietary additives in a buffer optimized for PCR.
How does SYBR Green work?
The SYBR green in the sample binds to the herring sperm DNA and, once bound, fluoresces giving off green light when illuminated by blue light. SYBR Green finds usage in several areas of biochemistry and molecular biology. It is used as a dye for the quantification of double stranded DNA in some methods of quantitative PCR .
What is SYBR Green qPCR?
SYBR Green for qPCR. SYBR ® Green I is a commonly used fluorescent dye that binds double-stranded DNA molecules by intercalating between the DNA bases. It is used in quantitative PCR because the fluorescence can be measured at the end of each amplification cycle to determine, relatively or absolutely, how much DNA has been amplified.