Product Description:

SYBR Green I dye is a fluorescent dye that binds directly to double-stranded DNA(dsDNA) and is the most commonly used DNA-binding dye for fluorescence quantitative PCR. In quantitative PCR, SYBR Green I can fluoresce after non-specific binding with double-stranded DNA (dsDNA), so the purpose of detecting the amplification of PCR products can be achieved by detecting the fluorescence intensity of SYBR Green I in the reaction system. In the free state, SYBR Green I emits weak fluorescence. Once it binds to dsDNA, its fluorescence increases by 1000 times. The total fluorescence signal from a reaction is proportional to the amount of dsDNA present, and increases with the increase of amplified product. Therefore, by detecting the fluorescence signal intensity in the PCR reaction solution, the target gene can be accurately quantified, and the melting temperature of the amplified target DNA fragment can also be determined.

Instructions for use:

When used, the PCR reaction mixture was configured and 10000×SYBR Green I concentrated solution was added to the PCR reaction system so that the final concentration was 0.5× (adjusted between 0.2× and 1×). The above operations are recommended on ice.

Note: ① Please refer to the instructions for using DNA polymerase for preparation of reaction solution and PCR amplification conditions.

② For the use of the Realtime PCR amplification instrument, please refer to the instructions of each instrument.

Note:

Effect of concentration on fluorescence PCR results

The concentration of SYBR Green I is a key factor to ensure the success of fluorescence quantitative PCR experiments. If the concentration of SYBR Green I is too low, the change in the fluorescence signal will be reduced, which means that the sample with low copy may not be detectable; At high concentrations, the PCR reaction will be inhibited and the efficiency of PCR reaction will be reduced. Therefore, in general, when using SYBR Green I, the concentration should be optimized according to the actual situation, and the final concentration of the reaction is between 0.2× and 1×.

Effect of magnesium ion concentration

The inhibitory effect of SYBR Green I on PCR reaction could be reduced by increasing the concentration of magnesium ion. We recommend that the concentration of magnesium ions in the fluorescent PCR reaction with SYBR Green I be 0.5 ~ 3mM higher than that in the ordinary PCR reaction without SYBR Green I."

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.