20×Sybr Green I(qPCR Grade)
The fluorescence signal of SYBR Green I combined with dsDNA can be enhanced by 800-1000 times. In the PCR reaction system, when an excessive amount of SYBR Green I fluorescent dye is added, it is specifically incorporated into the DNA double strand to enhance the fluorescence signal, while the fluorescence of SYBR Green I dye molecules that are not incorporated into the strand remains unchanged, ensuring that the increase in fluorescence signal is completely synchronized with the increase in PCR products. Fluorescence can be measured during annealing or elongation stages.
Usage: Apply 20 × Add SYBR Green I concentrate to the PCR reaction system to achieve a final concentration of 0.5 × (0.2 × To 1 × Adjust between).
Precautions:
1. The effect of concentration on fluorescence PCR results
The concentration of SYBR Green I is a crucial factor in ensuring the success of fluorescence quantitative PCR experiments. If the concentration of SYBR Green I is too low, it will reduce the changes in the fluorescence signal, which means that low copy samples may not be detectable. At high concentrations, it will inhibit the PCR reaction and reduce its efficiency. So generally, when using SYBR Green I, the concentration should be optimized according to the actual situation, and the final concentration of the reaction is 0.2 × To 1 × Between.
2. The effect of magnesium ion concentration
Increasing the concentration of magnesium ions can reduce the inhibitory effect of SYBR Green I on PCR reactions. We suggest that when using SYBR Green I for fluorescence PCR reaction, the magnesium ion concentration is 0.5-3mM higher than that of a regular PCR reaction without SYBR Green I.
Storage conditions: -20 ℃, avoid repeated freeze-thaw.
