Sahara Multiplex qPCR Master Mix

Highlights

• Multiplex up to four Real-Time PCR reactions with no effect on C_q’s
• Extreme thermal stability: withstands three months at 25 ºC with no impact on efficiency
• Overcomes inhibition in crude extracts and environmental samples
• Universal mix runs fast protocols, amplifies GC-rich targets, and incorporates aptamer-based hot start and dUTP carry-over prevention systems
• License-free for commercial or diagnostic use

Description

Sahara Multiplex qPCR Master Mix is a 2X Taq mix for probe-based Real-Time PCR. It quantitatively amplifies singleplex and multiplex (up to 4-plex) qPCR targets at high efficiency from purified samples and crude extracts.

The Most Stable Master Mix on the Planet

Stable at 50 ºC for eight days, or 25 ºC for three months, Sahara mixes ship economically at ambient temperatures, prevent C_q drift over time due to thermal degradation, and are ideal for high throughput or field applications where temperature control is not always ideal.

Amplifies Everything Without Inhibition

Sahara Multiplex qPCR Master Mix has been validated against genomic, plasmid, and cDNA targets from a wide range of organisms. The mix supports both standard and fast protocols, and amplifies GC-rich targets up to 70%.

Numerous enhancers bind & inactivate PCR inhibitors, allowing amplification from crude extracts of:

• Blood
• Food & beverages
• Plant & animal tissue
• Environmental air & water samples

Multiplex with Confidence

Multiplex qPCR often suffers from competitive inhibition, where a high copy number target amplifies early, consuming reactants that prevent lower copy number targets from amplifying. With high levels of Taq, dNTPs, and multiplexing enhancers, Sahara Multiplex has been demonstrated to amplify four multiplex targets simultaneously, with no change in C_q when compared with singleplex reactions.

Simplify your PCR

Set up reactions with ease at room temperature: an aptamer-based hotstart system prevents amplification from occurring below 50 ºC, while a blue non-interfering loading dye helps avoid pipetting errors. When activated by the addition of UNG, a dUTP carry-over prevention system prevents false positives from amplicon contamination.

Performance

Exceptional Linearity & Dynamic Range

Ara h2 gene fragment

E = 99.96%, R² = 1.000

Dynamic range of 10¹⁰ is demonstrated while maintaining excellent efficiency and linearity using Sahara Multiplex qPCR Master Mix to amplify a 157 bp gene fragment on an Open qPCR instrument.

High Efficiency, Even When Fourplexing

Four targets were amplified simultaneously using Sahara Multiplex qPCR Master Mix on a Bio-Rad CFX Touch instrument. 10-fold dilutions of all targets achieved excellent linearity & efficiency.

Get the Same C_q, Whether Singleplex or Fourplex

When singleplex and fourplex reactions are compared, minimal shift in C_q values are observed for all dilutions of targets.

Incredibly Stable

Sahara Multiplex qPCR Master Mix was stored at 50 ºC, and fourplex reactions were run daily on Bio-Rad CFX96 Touch instrument. Consistent C_q values were observed throughout the trial.

Fast & Quantitative

Lambda gDNA, 72 bp amplicon

E = 97%, R² = 0.996

A fast 19 minute protocol was run on an Open qPCR instrument, obtaining excellent linearity & efficiency. Protocol: Initial denaturing 30s @ 95 ºC, Cycle 30x: 3s @ 95 ºC, 14s @ 60 ºC, 5 C ºC/s ramp times.

Fast & Effective Hot Start

Hot Start

No Hot Start

At left, a challenging 67% GC Salmon sperm assay was run with and without Sahara Multiplex qPCR Master Mix’s hot start aptamer. The aptamer hot start successfully prevents the non-specific products seen in the non-hot start assay. The extra small band in the hot start formulation is the disassociated aptamer.

A challenging 67% GC Salmon sperm assay was run with and without Sahara Multiplex qPCR Master Mix’s hot start aptamer. The aptamer hot start successfully prevents the non-specific products seen in the non-hot start assay. The extra small band in the hot start formulation is the disassociated aptamer.

Hot Start

No Hot Start

At right, a duplex assay targeting Enterococcus gDNA (pink) and Yeast gDNA (blue) was run with and without the aptamer hot start. The assay lacking hot start produced inconsistent plateaus and poor curves due to non-specific amplification. The hot start assay produces consistent plateaus for high and low dilutions of target.

A duplex assay targeting Enterococcus gDNA (pink) and Yeast gDNA (blue) was run with and without the aptamer hot start. The assay lacking hot start produced inconsistent plateaus and poor curves due to non-specific amplification. The hot start assay produces consistent plateaus for high and low dilutions of target.

Specifications

PCR

^† user must add UNG to enable

Shipping, Storage, and Stability

Compatible Instruments

The mix ships with an optional ROX reference dye, allowing it to be used with high, low, and no-ROX qPCR instruments. 

FAQs

• What is a PCR Master Mix?

  A PCR master mix is a premixed solution that contains most of the components necessary to run a PCR assay. The mix contains Taq DNA polymerase, dNTPs, MgCl2, as well as enhancers and stabilizers in a buffer that is optimized for DNA amplification by PCR. Read more...

License

To promote collaboration in science, Sahara Multiplex qPCR Master Mix is provided under an open license:

• It may be used freely for any application, including commercial and diagnostic use
• No license agreements or royalty payments required for OEM use
• May be incorporated into your own test kits without restriction

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