Is SYBR Green an intercalating dye?

SYBR® Green is a dsDNA-binding dye that intercalates nonspecifically into dsDNA, allowing measurement of the amount of PCR product. As amplification proceeds, the amount of DNA product increases, and hence the number of SYBR® green molecules incorporated into DNA increases.

What makes SYBR Green dye a good intercalating dye?

SYBR Green is a green fluorescent cyanine dye that has high affinity for double-stranded DNA. The mode of binding is believed to be a combination of DNA intercalation and external binding.

What does intercalating dye mean?

In biochemistry, intercalation is the insertion of molecules between the planar bases of deoxyribonucleic acid (DNA). This process is used as a method for analyzing DNA and it is also the basis of certain kinds of poisoning.

How do I get rid of SYBR Green?

All Answers (9) DNA up to 100 mM NaCl and add 2½ volumes of absolute or 95% ethanol. After incubating for about 20 minutes on ice, centrifuge mixture for at least 10 minutes in a microfuge at 4°C. Remove the ethanol and wash the pellet once with 70% ethanol.

Is SYBR Green used in qPCR?

SYBR Green is one of the most commonly used fluorescent dyes in qPCR. Therefore, the fluorescence intensity is directly associated with the concentration of double-stranded DNA, which can be measured at the end of each amplification cycle to determine the PCR progress.

Is SYBR Green Green?

The SYBR family of dyes is produced by Molecular Probes Inc., now owned by Thermo Fisher Scientific. SYBR Green I binds to DNA. The resulting DNA-dye-complex best absorbs 497 nanometer blue light (λmax = 497 nm) and emits green light (λmax = 520 nm).

Why are intercalating agents bad?

A substance that inserts itself into the DNA structure of a cell and binds to the DNA. This causes DNA damage. In cancer treatment, DNA intercalating agents may kill cancer cells by damaging their DNA and stopping them from dividing.

What do intercalating agents do to DNA?

Intercalating Agents Insertion of these agents distorts the DNA double helix, thereby interfering with DNA replication, transcription, and repair. Such DNA distortions often result in mutations, so intercalating agents are also mutagens.

Is SYBR Safe an intercalating dye?

SYBR Safe is one of a number of SYBR dyes made by the Life Technologies Corporation. SYBR Safe binds to DNA. The resulting DNA-dye-complex absorbs blue light (λmax = 509 nm) and emits green light (λmax = 524 nm)….SYBR Safe.

Is SYBR Green Safe?

SYBR Safe DNA Gel Stain was specifically developed as a safer alternative to ethidium bromide. Invitrogen SYBR Green I Nucleic Acid Gel Stain is an ultrasensitive stain for dsDNA, and Invitrogen SYBR Green II RNA Gel Stain is a highly sensitive stain for RNA and ssDNA.

How does qPCR work with SYBR Green?

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.

Are there any alternatives to SYBR Green I?

This is surprising considering the reported limitations of SYBR Green I, which include limited dye stability, dye-dependent PCR inhibition, and selective detection of amplicons during DNA melting curve analysis of multiplex PCRs. We have tested an alternative to SYBR Green I and report the first detailed evaluation of the intercalating dye SYTO9.

When was SYBR Green I used for PCR?

Following the initial report of the use of SYBR Green I for real-time polymerase chain reaction (PCR) in 1997, little attention has been given to the development of alternative intercalating dyes for this application. This is surprising considering the reported limitations of SYBR Green I, which inc …

What is the concentration of SYBR Green I?

The concentrations of the dyes (Invitrogen, CA, USA) used in this study ranged from 0.1 to 20 μM. The molecular concentration of SYBR Green I was calculated using the value from Zipper et al. [28]. It was estimated that a 1× SYBR Green I solution was equivalent to 2 μM.

How is SYBR Green similar to ethidium bromide?

SYBR green is an intercalating dye and functions similar to ethidium bromide (Figure 8). SYBR green binds more efficiently to double stranded (deoxyribonucleic acid) The molecule that encodes genetic information.