SYBR^®Green Based Quantitative PCR

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.

Russell Higuchi and his coauthors recognized in 1993 that PCR could be monitored in real time by using fluorescent labels bound to the accumulating PCR product. SYBR^® Green I is one such fluorescent DNA-binding dye that can be included in PCR buffer. When bound to DNA, the dye changes structurally and becomes less mobile, causing its energy to be released as fluorescence. The fluorescence increases along with the concentration of DNA.

SYBR® Green I has an excitation and emission maxima of 494 nm and 521 nm, respectively.

All of our SYBR^® Green products are formulated with a hot-start mechanism for enhanced accuracy, precision, and sensitivity. For speed and efficiency, both LuminoCt^® and KiCqStart^® product lines are designed for Fast qPCR; the JumpStart™ products provide equally quality reagents for conventional qPCR.

Fast/Conventional qPCR:

• KiCqStart – Formats for all major qPCR instruments
• LuminoCt – Compatible with most cyclers

Conventional qPCR

• JumpStart

KiCqStart SYBR^® Green ReadyMix™ – Optimized for Bio-Rad, Qiagen, Instruments

KiCqStart products are designed with a stringent hot-start mechanism for greater specificity. They are prepared as ReadyMixes with non-custom reaction components for convenience. They are optimized for fast or conventional qPCR and are formatted with varying levels of ROX, the passive reference dye, at concentrations recommended by cycler manufacturers. Find the right KiCqStart product for your instrument and pair it with a Primer and Primer Gene Array.

Full Instrument Compatibility Chart

LuminoCt ReadyMixes – Highly Optimized

LuminoCt performs fast qPCR in as little as 25 minutes without sacrificing accuracy, precision, or sensitivity. It is produced as a highly optimized ReadyMix developed with JumpStart Taq antibody for rapid activation and unparalleled sensitivity. The ReadyMix virtually eliminates the need to optimize assay conditions and is compatible with commonly available qPCR instruments. Often users are able to simply purchase the kit, build their assay, and get right to work.

Conventional qPCR

JumpStart Taq ReadyMix for Conventional qPCR

SYBR^® Green JumpStart Taq Readymixes provide a convenient solution for conventional qPCR experiments. The JumpStart Taq antibody delivers antibody-inactivated hot-start PCR which prevents non-specific product formation. The product line features options for optimization and offer compatibility with tube-, plate-, and capillary-based instruments.

Figure 1. Linearity of pBAC-2cp 10-fold dilutions with SYBR Green JumpStart Taq ReadyMix

Figure 2. Ct values for Λ Amplicon using SYBR Green JumpStart Taq ReadyMix