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All-in-One™ qPCR Mix and Validated Primers — universal reaction conditions for all qPCR primers

20% Off qPCR Kits

 

Introduction

All-in-One™ qPCR mix with validated primers provide universal qPCR reaction conditions and robust quantitative PCR data. The jointly developed and co-optimized All-in-One qPCR mix and gene-specific primers deliver the entire range of advantages you need without any of the high costs you’ve come to expect:

  • Uniform reaction conditions reduce experimental design time enabling earlier journal submissions
  • High amplification efficiency and sensitivity even for low-copy genes means reliable quantitation every time
  • Absence of non-specific amplification* and no primer-dimers* ensures reproducible and ready-to-publish data

The All-in-One qPCR mix uses high-fidelity hot-start polymerase, an optimized reaction buffer and high-quality dNTPs to enable specific and sensitive amplification from even low-copy RNA (cDNA) or DNA species (Figure 1).

 

Figure 1. qPCR reaction using the All-in-one qPCR Master Mix. One microgram of total RNA from MCF-7 cells was reverse transcribed using GeneCopoeia  All-in-One cDNA synthesize kit. The RT reactions were diluted to 1pg to 1ng of RNA /µl. 1 µl of the diluted  RT reactions were used in the subsequent qPCR for 35 cycles using GeneCopoeia All-in-One qPCR Mix (A).  The end products were run on 2% Agarose gel (B).

 

 

Amplification curves Melting Curves Standard Curve

 

Figure 2. The amplification efficiency and detection sensitivity of the 2X All-in-One™ qPCR Mix are assessed by standard curves made by gradient dilution of plasmid DNA from 5×106 to 5 molecules. The peak values from amplification and melting curves show that very high sensitivity can be obtained using All-in-One™ qPCR Mix which can detect as low as 5 molecules. At the same time, high amplification efficiency has also been shown by a good linear relationship among each concentration.

 

All-in-One qPCR validated primers get the job done     

Eliminate endless adjustments and optimizations with precision qPCR from GeneCopoeia. Just add All-in-One primers and start.

The All-in-One qPCR human- mouse- or rat-specific primers are designed by a proprietary algorithm and validated for precision performance. Primer validation includes melting curve to ensure amplification of the correct target DNA (figure 2).

When used in combination with the All-in-One SYBR® Green qPCR Mix, the All-in-One primers deliver reliable and reproducible high performance in quantitative PCR assays.

Amplification curves
Melting Curves Agarose Gel Electrophoresis Validation Result

 

Figure 3. Forty-five pairs of gene-specific All-in-One™ qPCR primers were experimentally validated to yield a single dissociation curve peak and to generate a single amplification of the correct size for the targeted genes. A cDNA pool, containing reverse transcribed products of total RNA from 10 different human tissue (lung, liver, testicle, ovary, spleen, brain, placenta, pancreas, heart and mammary), was used as the qPCR validation template. qPCR was performed using 0.2 µM primer with 2X All-in-One qPCR Mix. Reactions were incubated for 10 min. at 95°C, followed by 40 cycles of 95°C for 10 sec.; 60°C, 20 sec. and 72°C, 15 sec. using Bio-Rad iQ5 Instrument. At the end of the last cycle, the temperature was increased from 72 to 95°C to produce a melting curve. Amplification curves, melting curves and agarose gel electrophoresis (validation result for positive in odd-lane and no template control (NTC) in even lane) shown for the 10 samples.

* Non-specific amplification and absence of primer-dimers are ensured when All-in-One validated PCR primers and PCR mix are used together.
† Primers validated automatically for human, mouse and rat. Inquire for validation of other species.

qPCR primers and kits

Products

 
Buy Catalog# Product Description Price
QP001 All-in-One™ qPCR Mix (200 qPCR reactions) High-fidelity, hot-start DNA polymerase,ROX reference dye,optimized reaction buffer and dNTPs $169
QP002 All-in-One™ qPCR Mix (600 qPCR reactions) High-fidelity, hot-start DNA polymerase,ROX reference dye,optimized reaction buffer and dNTPs $389
QP004 All-in-One™ qPCR Mix (1000 qPCR reactions) High-fidelity, hot-start DNA polymerase, ROX reference dye, optimized reaction buffer and dNTPs $635
QP005 All-in-One™ qPCR Mix (4000 qPCR reactions) High-fidelity, hot-start DNA polymerase, ROX reference dye, optimized reaction buffer and dNTPs $1995
QP008 All-in-One™ First-Strand cDNA Synthesis Kit for gene qPCR array (20 RT reactions) M-MLV RT (RNase H-), Reaction Buffer, RNase Inhibitor, dNTPs, Oligo(dT)18, Random Primer, dd H2O and spike-in control (for use with gene qPCR arrays) $129
QP009 All-in-One™ First-Strand cDNA Synthesis Kit for gene qPCR array (60 synthesis reactions) M-MLV RT (RNase H-), Reaction Buffer, RNase Inhibitor, dNTPs, Oligo(dT)18, Random Primer, dd H2O and spike-in control (for use with gene qPCR arrays) $285
  Variable All-in-One™ qPCR Primer
(200 reactions)
Validated gene-specific for human, mouse and rat toensure specificity and sensitivity

 

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Publications

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