Whip your experiments into shape with the toughest mix around
Experience the Amplification Difference. ToughMix®.
Quantabio ToughMix chemistry is the trade secret difference to our mastermix formulations. It has helped scientists for more than 20 years to amplify and analyze challenging samples by overcoming common PCR inhibitors. Widely used across a multitude of molecular biology applications, ToughMix formulations can be used directly from crude lysates as well as following an extraction protocol.
What can ToughMix do for you
- Work directly with crude lysates
- Avoid expensive and time-consuming purification steps
- Compatible with a wide range of probe designs and detection chemistries
- High quality Taq polymerase – free of residual host DNA
- Neutralize problem causing inhibitors present in crude samples
Inhibitors | Common Source | Reagent performance | |
Competitor | ToughMix | ||
Polyphenols | Plant extracts | – | |
Humic acid | Soil Plant tissues |
– | |
Hematin | Dried bloods Blood spots |
– | |
Hemoglobin | Blood | ||
Polysaccharides | Feces Plant tissues |
– | |
Melanin | Hair Skin |
– |
Inhibitor Resistance of ToughMixes
High sensitivity in the presence of inhibitors. Comparison of PerfeCTa® qPCR ToughMix sensitivity against other enzyme manufacturers in the presence of hemoglobin (left panel) and humic acid (right panel). ToughMix maintained Cq values compared to control experiments when challenged with inhibitors.
Robust PCR Amplication with Increasing Concentration of Inhibitors
Inhibitor Resistance of AccuStart II PCR ToughMix against different PCR inhibitors. A 1-kb fragment from 1 x 104 copies of the Tetracyclin resistance gene was amplified in 20 μL reaction volumes according to the recommended protocol. Reactions were challenged with varying concentrations of different PCR inhibitors and analysed by gel electrophoresis.
Hemoglobin: 316 – 3.16 ng/μL, Humic Acid: 31.6 – 0.316 ng/μL,
Hematin: 100 – 1 μM, Melanin: 10 – 0.1 ng/μL,
C: control reactions without inhibitor, L: 1 Kb Plus DNA Ladder.
Hemoglobin: 316 – 3.16 ng/μL, Humic Acid: 31.6 – 0.316 ng/μL,
Hematin: 100 – 1 μM, Melanin: 10 – 0.1 ng/μL,
C: control reactions without inhibitor, L: 1 Kb Plus DNA Ladder.
Inhibitor Resistance for 1-Step RT-qPCR ToughMix
Inhibitor resistance for 1-Step RT-qPCR ToughMix. c-myc was amplified from 10 ng total RNA using qScript XLT 1-step RT-qPCR ToughMix in the presence of increasing concentrations of hemoglobin. Ct values are maintained even when challenged with up to 400 ng/μL inhibitor.
Resistance to a Wide Range of Inhibitors
Resistance to a wide range of inhibitors. A 2 kb λ DNA template was amplified using each manufacturers recommended cycling conditions with different amounts of inhibitors. The experiment was run in duplicate.
ToughMix technology is available for different applications:
Conventional PCR
RT-PCR
SYBR-based qPCR
Probe-based qPCR
RT-qPCR
Features
Concentration
2x
2x
Kit Format
1 tube
1 tube mix + 1 tube loading dye
Amplicon Length
Up to 24kb
Up to 4kb
Applications
High fidelity PCR, Crude PCR, Fast PCR, Long PCR, High Yield, NGS
Routine PCR, Crude PCR
Available Formats
Master Mix
Packaging (Rxns/Units)
25 Rxns
100 Rxns
500 Rxns
100 Rxns
500 Rxns
100 Rxns
800 Rxns
4000 Rxns
800 Rxns
4000 Rxns