Traditional QPCR

Probe Based Quantitative PCR

Probe based QPCR relies on the sequence-specific detection of a desired PCR product. Unlike SYBR® based QPCR methods that detect all double-stranded DNA, probe based QPCR utilizes a fluorescent-labeled target-specific probe resulting in increased specificity and sensitivity. Additionally, a variety of fluorescent dyes are available so that multiple primers can be used to simultaneously amplify many sequences.

PCR Technical Animations

• Ideal for high throughput. ReadyMixes contain all the necessary components for QPCR; you simply add the fluorescent detection chemistry, primers, and template.
• Specific. JumpStart Taq antibody prevents non-specific product formation through its hot-start mechanisms.
• Maximum flexibility in detection method since no detection method has been incorporated into the formulation.
• Optimized formulations with the addition of dUTP to facilitate carry–over prevention from previous PCR.

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Description

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OLIGO Custom DNA and RNA Oligos Design and Order Custom DNA Oligos
PROBES Custom qPCR Probes Design and Order DNA, LNA, and BHQ Probes for qPCR
D6442 JumpStart Taq ReadyMix for High Throughput Quantitative PCR Ready-to-use 2x mix for qPCR with ROX
D7440 JumpStart Taq ReadyMix for Quantitative PCR For probe-based real-time PCR
L6669 LuminoCt® qPCR ReadyMix For fast probe-based quantitative PCR

Quantitative RT-PCR

Product #

Description

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OLIGO Custom DNA and RNA Oligos Design and Order Custom DNA Oligos
PROBES Custom qPCR Probes Design and Order DNA, LNA, and BHQ Probes for qPCR
QR0200 Quantitative RT-PCR ReadyMix One step RT-qPCR for probe-based methods, MMLV & hot-start Taq
QR0100 SYBR® Green Quantitative RT-qPCR Kit One step SYBR® Green RT-qPCR with MMLV & hot-start Taq DNA Polymerase

Reagents

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R4526 Reference Dye for Quantitative PCR 100 ×, solution

SYBR® Green Based Quantitative PCR

SYBR® Green I, a commonly used fluorescent DNA binding dye, binds all double-stranded DNA and detection is monitored by measuring the increase in fluorescence throughout the cycle. SYBR Green I has an excitation and emission maxima of 494 nm and 521 nm, respectively. Specificity of Sigma’s SYBR based QPCR detection is greatly enhanced by the incorporation of a hot-start mediated Taq polymerase, JumpStart Taq.
SYBR Green I Based QPCR Technical Animation

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Description

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KSPQ12012 KiCqStart® SYBR® Green Primers Predesigned primers for gene expression analysis
KSGA2013 KiCqStart® SYBR® Green Primers Gene Arrays Primer arrays for expression analysis of gene families
KCQS00 KiCqStart® SYBR® Green qPCR ReadyMix For Bio-Rad, Cepheid, Eppendorf, Illumina, Corbett, and Roche systems
KCQS01 KiCqStart® SYBR® Green qPCR ReadyMix Low ROX, for ABI and Stratagene instruments
KCQS02 KiCqStart® SYBR® Green qPCR ReadyMix with ROX for ABI instruments
KCQS03 KiCqStart® SYBR® Green qPCR ReadyMix iQ, with fluorescein for Bio-Rad systems
S4438 SYBR® Green JumpStart Taq ReadyMix for quantitative PCR, MgCI2 in buffer
S5193 SYBR® Green JumpStart Taq ReadyMix for quantitative PCR, without MgCl2 in buffer
S9194 SYBR® Green JumpStart Taq ReadyMix for High Throughput qPCR SYBR® Green qPCR reagent, passive reference dye included
S1816 SYBR® Green JumpStart Taq ReadyMix for Quantitative PCR, Capillary Formulation SYBR® Green qPCR reagent for Roche LightCycler® capillary systems