RT-qPCR SYBR Green Mixes
SYBR Green is an aromatic organic compound of chemical formula C32H37N4S belonging to the asymmetric cyanines (fluorophores). It has the ability to bind to nucleic acids and emit fluorescence. It is therefore used in molecular biology as a fluorescent agent, especially when carrying out quantitative PCRs.
SYBR Green is the most commonly used dye for non-specific detection. SYBR green I is a molecule that can bind to all types of double stranded nucleic acids. It is not an intercalating agent: by definition, an intercalating agent is a molecule that can be inserted between the trays formed by the paired bases of a nucleic acid; The SYBR green does not meet this definition. Once fixed, it becomes a very good fluorophore. Thus, the double-stranded DNA / SYBR green I complex absorbs blue light (λmax = 497 nm) and emits green light (λmax = 520 nm). The emitted fluorescence can be detected and related to the amount of target.
Some real-time PCR instruments require an internal reference dye for fluorescent signal normalization and correction of optical well-to-well variations. The variation in fluorescence may result from the design of the instrument. The ROX Passive Reference Dye is an inert additive that provides a constant fluorescent signal for standardization of the sample throughout the qPCR test in real time.
CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon®, Opticon 2,
Cepheid SmartCycler®; Eppendorf Mastercycler® ep realplex, realplex 2 s;
Illumina Eco qPCR;
Qiagen/Corbett Rotor-Gene® Q, Rotor-Gene® 3000, Rotor-Gene® 6000;
Roche Applied Science LightCyclerTM 480;
Thermo Scientific PikoReal Cycler.
Biosystems 7500, 7500 Fast, ViiA7; Stratagene MX4000,
Biosystems 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast;