QuantiNova Probe RT-PCR Kit (500)，208354，Qiagen，凯杰

QuantiNova RT-PCR Kits

For highly sensitive and specific one-step qRT-PCR and multiplex qRT-PCR using sequence-specific probes or one-step qPCR using SYBR® Green I for gene expression analysis

Unique two-phase hot-start procedure for room-temperature setup

Visual pipetting control, resulting in fewer pipetting errors

Internal control for positive in-process verification of successful RT-PCR

Accurate quantification over several logs of template

Sensitive detection of up to 5 targets in 1 tube

Principle of the novel QuantiNova two-phase hot-start mechanism.

At ambient temperature the HotStaRTScript is inhibited by the RT-Blocker and the QuantiNova DNA Polymerase is kept inactive by QuantiNova Antibody and QuantiNova Guard. At 50°C the RT is activated while the QuantiNova DNA polymerase remains inactive. At 95°C the RT enzyme is denatured and the DNA polymerase is activated.

Accurate reaction setup indicated by the built-in pipetting control.

The master mix contains an inert blue dye. Combined with QuantiNova Yellow Template Dilution Buffer, the resulting solution turns green, indicating that the reaction was set up correctly.

Convenient room-temperature reaction setup without compromising performance.

Real-time RT-PCRs including master mix, template RNA, primers and probes for [A] GAPDH, [B] MYC and [C] HSP90AA1 were left at room temperature (22°C) for 1 h and 2 h. These samples were run in parallel to freshly prepared reactions (set up on ice) on the Bio-Rad CFX 384 instrument. Duplicate reactions of 3 different template amounts were tested. The Cq values provided by the QuantiNova Probe RT-PCR Kit did not vary after extended storage of the samples at room temperature.

Increased reliability of gene expression results using gDNA reduction.

Total HeLa RNA (10 ng/rnx) samples were spiked with increasing amounts of human genomic DNA. Expression of MGAT1 was analyzed by qRT-PCR. MGAT1 is a single-exon gene that does not allow the discrimination of RNA and gDNA by assay design. Without the gDNA reduction step, Cq values decreased linearly, falsely indicating an increase in expression rate. In contrast, the expression levels remained similar when using the gDNA reduction step.

QuantiNova RT-PCR Kits (real-time RT-PCR kits) enable sensitive quantification of RNA targets by real-time one-step PCR using SYBR Green I or sequence-specific probes. A combination of various integrated safety features removes variables and prevents artifacts, ensuring reliable gene expression profiling. The combination of a unique two-phase hot-start and PCR buffer system in the ready-to-use master mix allows room-temperature setup and ensures highly sensitive qRT-PCR on any real-time cycler. The optional QuantiNova Internal Control RNA can be used to test successful reverse transcription and amplification. It is intended to report instrument or chemistry failures, errors in assay setup and the presence of inhibitors. Furthermore, the visual pipetting control can be used to monitor correct pipetting. The integrated gDNA reduction step in the QuantiNova Probe RT-PCR procedure prevents overquantification of transcripts caused by genomic DNA carryover.

The QuantiNova Multiplex RT-PCR Kit enables fast and reliable quantification of up to 5 RNA targets in a single tube by multiplex real-time RT-PCR. Our Q-Bond technology and optimized master mix promote ultrafast multiplex real-time RT-PCR within 1 hour. The combination of a unique two-phase hot-start procedure with our multiplex PCR buffer system ensures highly sensitive qRT-PCR on any real-time cycler without the need for optimization, and enables automated reaction setup at room temperature. The kit also provides protocols for extracted RNA and direct amplification from cultured cells, even down to a single cell. The optional QuantiNova Internal Control RNA can be used to monitor successful reverse transcription and amplification, and the visual pipetting control can be used to monitor correct pipetting.

The QuantiNova SYBR Green and Probe RT-PCR Kits can be used for gene expression analysis of RNA targets on any real-time cycler. This includes the Rotor-Gene Q and instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche and Agilent.

The QuantiNova Multiplex RT-PCR Kit can be used for multiplex gene expression analysis of RNA targets on any real-time cycler. To fully benefit from multiplexing, we recommend using an instrument that provides up to 5-plex capacity, such as the Rotor-Gene Q.