QuantiTect Probe PCR Kit (200)

• Highly sensitive detection of low-copy targets
• qPCR using sequence-specific probes or SYBR Green
• Accurate quantification over several logs of template
• No need to optimize reaction and cycling conditions
• Detection of up to 5 targets in the same tube

QuantiTect PCR Kits (qPCR kits) enable sensitive quantification of gDNA and cDNA targets by real-time PCR and two-step RT-PCR using sequence-specific probes or SYBR Green I detection. The real-time PCR kits also allow reliable quantification of up to 5 gDNA or cDNA targets in a single tube by multiplex, real-time PCR or two-step RT-PCR. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive qPCR on any real-time cycler without the need for optimization. The dNTP mix includes dUTP, allowing optional treatment with UNG. For convenience, the master mixes in QuantiTect PCR Kits can be stored at 2–8°C.

Two kit formats are available for multiplex PCR using sequence-specific probes: the QuantiTect Multiplex PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiTect Multiplex PCR NoROX Kit for all other cyclers.

HotStarTaq DNA Polymerase included in QuantiTect quantitative PCR kits increases the specificity of the PCR reaction by providing the most stringent hot start compared with other polymerases.

QuantiTect SYBR Green PCR Kits allow specific quantification over a wide linear range (see figure " Specific quantification over a wide linear range"). When used in combination with the QuantiTect Reverse Transcription Kit and QuantiTect Primer Assays, QuantiTect SYBR Green PCR Kits deliver sensitive and reliable results (see figure " Specific and sensitive quantification").

QuantiTect Probe PCR Kits deliver sensitive and reliable results in combination with the QuantiTect Reverse Transcription Kit (see figure " High sensitivity and efficiency, and wide dynamic range"). The unique PCR buffer composition enables QuantiTect Probe PCR Kits to provide sensitive quantification of low-copy DNA targets, as well as accurate quantification over a wide linear range (see figure " Wide dynamic range in real-time PCR").

The QuantiTect Multiplex PCR Master Mix ensures that the PCR products in a multiplex reaction are amplified with the same efficiency and sensitivity as the PCR products in the corresponding single-amplification reactions (see table “Comparable threshold cycle (C_Τ) values with triplex and singleplex PCRs”, and figure “ Comparable results with 4-plex and singleplex PCRs”.

As few as 10 copies of a target gene can be detected with the kits, even if the copy number of the reference gene in the same reaction is up to 10⁶-fold greater (see figure “ Detection of 10 copies of target gene with excess reference gene" and the corresponding table "Successful quantitative triplex qPCR of low and high abundance targets").

QuantiTect SYBR Green PCR Kits contain an optimized, ready-to-use master mix for highly specific and sensitive real-time quantification cDNA targets using SYBR Green I (see table “Components of 2x QuantiTect SYBR Green PCR Kit”). The fluorescent dye SYBR Green I in the master mix enables the analysis of many different targets without having to synthesize target-specific labeled probes. A balanced combination of K⁺ and NH₄⁺ ions in the PCR buffer promotes specific primer annealing and enables high PCR specificity and sensitivity (see figure "Specific primer annealing"). In addition, HotStarTaq DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.

QuantiTect Probe PCR Kits contain an optimized, ready-to-use master mix for highly specific and sensitive real-time quantification of gDNA and cDNA targets using sequence-specific probes (see table “Components of 2x QuantiTect Probe PCR Kit”). The kits are designed for use with all types of sequence-specific probes, including hydrolysis probe detection (e.g., TaqMan® and other dual-labeled probes), FRET probes, and Molecular Beacons. QuantiTect Probe PCR Kits contain a unique PCR buffer that contains a balanced combination of K⁺ and NH₄⁺ ions, which promote specific primer annealing, enabling high PCR specificity and sensitivity (see figure "Specific primer annealing"). In addition, HotStarTaq DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.

QuantiTect Multiplex PCR Kits enable success in multiplex, two-step RT-PCR on the first attempt (see flowchart " QIAGEN multiplex kits"). The optimized master mix ensures that PCR products in a multiplex reaction are amplified with the same efficiency and sensitivity as PCR products in a corresponding single-amplification reactions. As few as 10 copies of a target gene can be detected with the kit.

Amplifying reference and target genes in the same reaction instead of in separate reactions increases the reliability of gene quantification by minimizing handling errors. The QuantiTect Multiplex PCR Master Mix contains a balanced combination of K⁺ and NH₄⁺ ions as well as the unique synthetic Factor MP, which together promote stable and efficient annealing of primers and probes to the nucleic acid template, enabling high PCR efficiency (see table "Components of 2x QuantiTect Multiplex PCR Kit"). In addition, HotStarTaq DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.

The master mixes in QuantiTect PCR Kits also contain dUTP, enabling pretreatment with uracil-N-glycosylase (UNG) prior to starting PCR, which ensures that any c