Sahara Multiplex qPCR Master Mix
- Multiplex up to four Real-Time PCR reactions with no effect on Cq’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
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 8 days, or 25 ºC 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.
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:
- 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 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 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.
Exceptional Linearity & Dynamic Range
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
E = 101.8%, R2 = 0.999
E = 99.4%, R2 = 0.999
E = 100.7%, R2 = 1.000
E = 99.9%, R2 = 0.983
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
Avg ΔCq 0.25
Avg ΔCq 0.31
Avg ΔCq 0.17
Avg ΔCq 0.16
When singleplex and fourplex reactions are compared, minimal shift in Cq values are observed for all dilutions of targets.
Sahara Multiplex qPCR Master Mix was stored at 50 ºC, and a fourplex reactions were run daily on Bio-Rad CFX96 Touch instrument. No difference in Cq is observed throughout the trial.
Fast & Quantitative
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
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.
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.
|Hot Start Mechanism||Aptamer|
|GC Content||40 – 70%|
|Multiplex Targets||Up to 4|
|dUTP Carryover Prevention||Yes†|
|Visible Loading Dye||Yes|
|Exonuclease Activity||5' → 3'|
|Supported Probes||TaqMan/Hydrolysis, Molecular Beacon, Scorpions|
|Supported Templates||Genomic DNA, cDNA, Plasmid DNA|
|Recommended Reaction Volume||10 – 50 µL|
† user must add UNG to enable
Shipping, Storage, and Stability
|High Temperature Stability||8 days @ 50 ºC, 3 months @ 25 ºC|
|Freeze/Thaw Cycles||Up to 20|
|Storage Conditions||-20 ºC, or 4 ºC for 6 months. Protect from light|
The mix ships with an optional ROX reference dye, allowing it to be used with high, low, and no-ROX qPCR instruments.
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...
Sahara Multiplex qPCR Master Mix may be used license-free for commercial or diagnostic applications, and is available in bulk quantities for OEM use.