TaqDog 2X Green Master Mixes

Ready-to-use PCR premixes include loading dye

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Easy setup provides reliable amplification
TaqDog 2X Green Master Mixes provide everything needed to perform PCR. Just add template and primers and then start amplifying. These master mixes are based upon our popular TaqDog DNA Polymerases and includes nucleotides, buffer, and a brilliant green dye allowing for amplified samples to be directly loaded onto gels. Formulated for a 50 uL reaction size, its robust processivity makes smaller reaction volumes an economical option for labs looking to stretch their lab’s dollars. TaqDog master mixes come in two versions, premium and hot start. Our TaqDog Premium master mix is our most affordable option and provides great results in most cases. The TaqDog Hot Start master mix includes an aptamer-based hot start technology that promises better yields with no interference from primer-dimers.

TaqDog 2x Master Mix vs Manually Made Buffer

Enhanced to conquer GC-rich templates
Some PCR master mixes come with a separate tube of “PCR enhancer” for more difficult templates, especially those with increased G-C nucleotide content. The additional hydrogen bonding for G-C pairs makes PCR less efficient — often resulting in weak results. Both types of TaqDog 2x Green Master Mixes also include their own unique enhancer reagents that can be added when amplification efficiency needs to be improved.

Why TaqDog 2x Master Mix?

New Hot Start TaqDog leads to cleaner PCR
Hot start PCR has been around for over 20 years, and has been conclusively shown to reduce amplification artifacts caused by low-temperature “false”-priming. Many methods have been developed over this time, but TaqDog Hot Start’s aptamer technology combines their best features while eliminating their worst drawbacks. It’s based upon a DNA construct engineered to form a secondary hairpin structure that reversibly-binds Taq at room temperature. When bound, the polymerase activity is inhibited at temperature at or below 45 °C. This prevents the extension of incorrectly primed regions in a sample’s template DNA. At higher temperatures, the aptamer falls off, and only strongly binding, and specific, primers can then initiate polymerization. This results in final PCR products that are clear of extra, unwanted bands, including the notoriously aggravating primer-dimer bands.

TaqDog Hot Start aptamers are synthetically prepared and HPLC purified for both a precise and defined composition which greatly reduces lot-to-lot variation. This also avoids contamination with trace amounts of mammalian DNA or cellular contents that can be associated with antibody preps. TaqDog Hot Start aptamers are inert and immune to proteases. They contain a 3′-cap that prevents their amplification during PCR or even ability to be cloned. Interference of aptamers with downstream applications has not been observed. Moreover, DNA aptamers do not become irreversibly denatured over time (due to high temperatures), but in contrast, allow for more robust amplification and higher yields. Unlike antibody-mediated inhibition, aptamers are able to re-bind and inhibit our polymerase after the completion of a PCR protocol. This is useful in applications where baseline signal is critical and even modest post-PCR activity (for example during long long-temperature “soak” steps common to many protocols) can lead to artifacts in data.

Aptamer Hot Start Eliminates primer-dimer bands

It ain’t rocket science any more

PCR used to be considered an art, but in the 30 years since its general acceptance into scientific experimentation it’s become one of the best-characterized bio-enzymatic processes. To this end, we ask you this simple question: why pay prices that don’t reflect this fact? All of Bulldog Bio’s PCR products are priced fairly and promise satisfying results. It’s that simple.

Species of Origin Thermus aquaticus
Enzyme Type Thermostable DNA polymerase
Form Recombinant – Engineered
Part Numbers TDPM020, TDPM100, TDHM020, TDHM100
5’–3′ Exonuclease Activity Yes
“A” Overhang Yes
3’–5′ Exonuclease Activity No
Processivity Rate 1 Kbp / minute
Amplification Limit confirmed to 5 Kbp
Reaction Buffer 2x concentration,  includes dNTPs
Reaction Size 50 µL
Tube Size 1250 µL of 2x mastermix
Loading Dye Included, green in color, composed of blue (~3,500 bp) and yellow (~50 bp )
GC Enhancer Yes, included in separate tube
Storage up to 1 year at -20°C
Shipping Temperature On ice
DNase Free Yes
RNase Free Yes
Protease Free Yes
Mycoplasma gDNA Free Yes, measured at <10 copies per unit of enzyme
E. coli gDNA Reduced Yes, Measured at >10 copies per unit of enzyme
Hot Start Yes.  Hot Start version is aptamer based