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Sahara Multiplex qPCR Master Mix

Sahara Multiplex qPCR Master Mix

$0.00


Highlights


  • Multiplex up to four Real-Time PCR reactions with no effect on Cq’s
  • Extreme thermal stability: withstands three months at 25 °C 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

Description


Sahara Multiplex qPCR Master Mix is a 2X 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 °C for 8 days, or 25 °C for 3 months, Sahara mixes ship economically at ambient temperatures, prevent Cq drift over time due to thermal degradation, and are ideal for high throughput or field applications where temperature control is not always ideal.

Manual cover
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%.

Thanks to numerous enhancers which bind & inactivate inhibitors, the mix resists inhibition in crude extracts from:

  • Food & beverages
  • Blood
  • 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 Cq 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 °C, 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.

Specifications


PCR
Concentration2X
Hot Start MechanismAptamer
GC Content40 – 70%
Multiplex TargetsUp to 4
Reaction SpeedFast
dUTP Carryover PreventionYes
Visible Loading DyeYes
Exonuclease Activity5' → 3'
Supported ProbesTaqMan/Hydrolysis, Molecular Beacon, Scorpions
Supported TemplatesGenomic DNA, cDNA, Plasmid DNA
Recommended Reaction Volume10 – 50 µL

user must add UNG to enable

Shipping, Storage, and Stability
High Temperature Stability8 days @ 50 °C, 3 months @ 25 °C
Freeze/Thaw CyclesUp to 20
Shipping ConditionsAmbient
Storage Conditions-20 °C, or 4 °C for 6 months. Protect from light
Compatible Instruments

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

Performance


Exceptional Linearity & Dynamic Range
Ara h2 gene fragment
E = 99.96%, R2 = 1.000
Ara h2 gene fragment
Standard Curve

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
Enterococcus gDNA
mGAPDH gene fragment
Yeast gene fragment
mActin gene fragment

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 Cq, Whether Singleplex or Fourplex
Ara h2 gene fragment
Lambda gDNA
mActin gene fragment
Enterococcus gDNA
  • Singleplex
  • Fourplex

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

Incredibly Stable
Ara h2 gene fragment
Lambda gDNA
mActin gene fragment
Enterococcus gDNA

Sahara Multiplex qPCR Master Mix was stored at 50 °C, and a fourplex reactions were run daily on Bio-Rad CFX96 Touch instrument. No difference in Cq is observed throughout the trial.

Fast & Quantitative
Lambda gDNA, 72 bp amplicon
E = 97%, R2 = 0.996
Ara h2 gene fragment
Standard Curve

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

Fast & Effective Hot Start
Ara h2 gene fragment
Standard Curve
Standard Curve

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.

Standard Curve
Standard Curve

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 (top) produced inconsistent plateaus and poor curves due to non-specific amplification. The hot start assay (bottom) produces consistent plateaus for high and low dilutions of target.

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...

View all related knowledge-base articles.

License


Chai encourages commercial use of its products. Sahara Multiplex qPCR Master Mix may be used commercially license-free, and is available in bulk quantities for OEM applications.

Resources


Certificates of Analysis
Lot 7501697