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QuantiNova SYBR Green PCR Kit

For unparalleled results using SYBR Green-based qPCR
  • Unmatched specificity due to novel hot-start mechanism
  • Precise reaction setup with a built-in visual indicator
  • Accurate and robust detection of targets down to one copy
  • Magnified throughput and speed with ultrafast cycling
  • Seamless performance with QuantiTect Primer Assays
Featuring enhanced specificity, sensitivity, speed, and process safety, the QuantiNova SYBR Green PCR Kit sets new standards in SYBR Green-based qPCR. A novel hot-start mechanism significantly increases the specificity of real-time PCR and optimized chemistry produces accurate quantitative results from cDNA or gDNA on any real-time PCR cycler with detection of even a single target copy. Finally, a built-in visual indicator ensures correct pipetting.
Cat No./ID: 208052
QuantiNova SYBR Green PCR Kit (100)
$135.00
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For 100 x 20 µl reactions: 1 ml 2x QuantiNova SYBR Green PCR Master Mix, 500 µl QuantiNova Yellow Template Dilution Buffer, 250 µl QN ROX Reference Dye, 1.9 ml Water
Cat No./ID: 208054
QuantiNova SYBR Green PCR Kit (500)
$636.00
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For 500 x 20 µl reactions: 3 x 1.7 ml 2x QuantiNova SYBR Green PCR Master Mix, 500 µl QuantiNova Yellow Template Dilution Buffer, 1 ml QN ROX Reference Dye, 1.9 ml Water
Cat No./ID: 208056
QuantiNova SYBR Green PCR Kit (2500)
$2,442.00
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For 2500 x 20 µl reactions: 15 x 1.7 ml 2x QuantiNova SYBR Green PCR Master Mix, 5 x 500 µl QuantiNova Yellow Template Dilution Buffer, 5 X 1 ml QN ROX Reference Dye, 5 x 1.9 ml Water
The QuantiNova SYBR Green PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Maximum performance with and without QuantiNova Yellow Template Dilution Buffer.
EGFR was amplified from HeLa cDNA using a QuantiTect Primer Assay and the QuantiNova SYBR Green PCR Kit. The template DNA was diluted from 10 ng to 0.01 ng using QuantiNova Yellow Template Dilution Buffer or Tris buffer, and reactions were performed on the [A] Bio-Rad CFX96 and the [B] Applied Biosystems ViiA 7 instruments. Resulting CT values were comparable for both buffers. All reactions were performed in triplicate.
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Superior and flexible performance of the QuantiNova SYBR Green PCR Kit.
EGFR, ERBB2 and MYC were amplified from HeLa cDNA on the Bio-Rad CFX96, the Applied Biosystems ViiA 7, and the Applied Biosystems StepOnePlus cyclers. Template DNA was diluted in Tris buffer or in QuantiNova Yellow Template Dilution Buffer. Regardless of cycler, cycling conditions, or dilution buffer used, the QuantiNova SYBR Green PCR Kit generates comparable high-quality results.
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Robust and sensitive detection of single-copy targets.
[A] The single-copy gene IL1R2 was detected in 30, 3, 0.3, and 0.03 ng (10,000–10 target copies) of leukocyte genomic DNA using the QuantiNova SYBR Green PCR Kit to generate a calibration curve for CT value versus number of target copies. The correlation was highly linear. The calibration curve was then used to determine the actual number of target copies in 60 reactions set up to theoretically contain a single copy.  [B] Theoretical number of copies expected in each reaction was calculated using Poisson's equation and compared to the actual number of copies determined with the calibration curve. Calculated and experimental frequencies of detection were highly concordant, demonstrating high sensitivity and robustness.
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Accurate quantification over a wide dynamic range.
Ten-fold dilutions of HeLa cDNA ranging from 100 ng to 10 fg were quantified on the Applied Biosystems ViiA 7 instrument and the Bio-Rad CFX96 using an in-house assay for beta-actin. [A] The QuantiNova SYBR Green PCR Kit delivers accurate results over a wide dynamic range spanning 8 orders of magnitude. [B] This range is confirmed by the comparable amplification plots from both instruments.
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Consistent and sensitive results on a wide range of real-time PCR cyclers.
EGFR in HeLa cells was analyzed on various cyclers, fully exploiting the fast-cycling capabilities of each. The QuantiNova SYBR Green PCR Kit provides consistent sensitivity, reproducibility, and efficiency on all tested cyclers, despite varying formats, cycling conditions, and requirement for a passive dye.
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Superior results compared to similar SYBR Green PCR kits.
The performance of the QuantiNova SYBR Green PCR Kit was compared to [A] a SYBR Green PCR Kit from supplier B on a Bio-Rad CFX96 cycler and [B] a SYBR Green PCR Kit from supplier L on the Applied Biosystems ViiA 7 cycler. The QuantiNova SYBR Green PCR Kit provides significantly lower CT values, higher reproducibility, and higher reaction efficiency.
Performance
Accurate and robust detection of a single DNA molecule

The high sensitivity of the QuantiNova SYBR Green PCR Kit results in accurate and robust detection of even a single target copy (see figure Robust and sensitive detection of single-copy targets). This extreme sensitivity is achieved even under ultrafast cycling conditions. 

The QuantiNova SYBR Green PCR Kit can be used with a wide range of template amounts. Its performance has been tested for a dynamic range spanning 8 orders of magnitude (from 100 ng to 10 fg cDNA) and regardless of instrument used, quantification was accurate and robust (see figure Accurate quantification over a wide dynamic range).

Highly consistent results on a wide range of real-time cyclers

The stringent specificity and exceptional sensitivity of the QuantiNova chemistry produces optimal results on the QIAGEN Rotor-Gene Q, but can be used on any real-time PCR cycler, regardless of format, fast-cycling capacity, and need for a passive reference dye. ROX provided with the QuantiNova SYBR Green PCR Kit is simply added to the master mix if required. Amplification and quantification results using the Rotor-Gene Q, Agilent Technologies Mx3005P, Applied Biosystems 7900 HT Fast, ViiA 7, StepOnePlus, and 7500 Fast, Roche LightCycler 480, and Bio-Rad CFX96 are invariably robust and sensitive (see figure Consistent and sensitive results with a range of real-time PCR cyclers).

Unparalleled performance compared to equivalent SYBR Green PCR kits

The QuantiNova SYBR Green PCR Kit delivers superior results compared to other SYBR Green PCR kits (see figure Superior results compared to similar SYBR Green PCR kits). Regardless of cycling conditions or instrument, the QuantiNova chemistry demonstrably generates lower CT values, higher reproducibility, and higher reaction efficiency.

Built-in visual indicator for accurate reaction setup

The master mix supplied with the QuantiNova SYBR Green PCR Kit contains an inert blue dye that does not interfere with the real-time PCR, but increases visibility in the tube or well. The QuantiNova Yellow Template Dilution Buffer contains an inert yellow dye. When template nucleic acid is diluted in QuantiNova Yellow Template Dilution Buffer and added to the master mix, the color of the solution changes from blue to green (see figure Accurate reaction setup indicated by the built-in pipetting control), providing a visual indication that each reaction was set up correctly.

The QuantiNova Yellow Template Dilute Buffer provides peace of mind during reaction setup but is not required to obtain superior results with the QuantiNova SYBR Green PCR Kit (see figure Maximum performance with and without QuantiNova Yellow Template Dilution Buffer). The QuantiNova chemistry is designed to be flexible, so it can be incorporated into any established workflow. Thus, performance is optimal with your amplifcation target of interest, and your instrument and dilution buffer of choice (see figure Superior and flexible performance of QuantiNova SYBR Green PCR Kit).

Principle
The QuantiNova SYBR Green PCR Kit delivers cDNA or gDNA analysis with the highest specificity because of a novel, antibody-mediated, hot-start mechanism (see figure Principle of the novel QuantiNova hot-start mechanism). At low temperatures, QuantiNova DNA Polymerase is kept in an inactive state by the QuantiNova Antibody and the novel additive QuantiNova Guard, that stabilizes the complex. This improves the stringency of the hot-start and prevents extension of nonspecifically annealed primers and primer dimers. Within 2 minutes of raising the temperature to 95°C, the QuantiNova Antibody and QuantiNova Guard are denatured and the QuantiNova DNA Polymerase is activated, enabling amplification.

Additionally, the QuantiNova chemistry includes features that streamline your workflow without loss of PCR sensitivity and efficiency. Inert dyes in the master mix and in the QuantiNova Yellow Dilution Buffer serve as a visual indicator that each reaction has been set up correctly. The blue color of the master mix increases visibility of contents in the reaction vessel and turns green upon adding template nucleic acids diluted in the QuantiNova Yellow Dilution Buffer (see figure Accurate reaction setup indicated by the built-in pipetting control). The unique formulation of the PCR buffer included in the QuantiNova SYBR Green PCR Kit enhances the stringency of the hot-start mechanism and shortens cycling steps so your PCR is done faster and your throughput can be increased.
Procedure
The QuantiNova SYBR Green PCR Kit contains a ready-to-use master mix that eliminates the need for optimization of reaction and cycling conditions. Simply add template gDNA or cDNA and primers to the master mix and follow the protocol in the handbook to achieve rapid and reliable results on any real-time cycler. ROX passive reference dye is provided in a separate tube and the concentrations of the dye can be adjusted depending on the type of cycler used. Thus, the QuantiNova SYBR Green PCR Kit can be used on virtually any real-time cycler, regardless of format or cycling conditions.
Applications
The QuantiNova SYBR Green PCR Kit can be used for SYBR Green-based gene expression analysis of cDNA targets and quantitative gDNA analysis on any real-time cycler. This includes instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, LIFE Technologies, Roche, and Agilent, as well as the Rotor-Gene Q (see figure Consistent and sensitive results with a range of real-time PCR cyclers).

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Kit Handbooks (1)
For highly sensitive, ultrafast, quantitative real-time PCR and two-step RT-PCR using SYBR Green
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Quick-Start Protocols (1)
For highly sensitive and specific real-time qPCR and two-step RT-PCR using SYBR Green
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