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2x High-Fidelity PCR Master Mix

PCR PreMix, PCR MasterMix, PCR SuperMix, Taq Master Mix

Catalog No. Product Name Size List Price (US$) Quantity
MB119-S 2x Hot-Start High-Fidelity PCR Master Mix 100 x 20 ul reactions 80.00
MB119-L 2x Hot-Start High-Fidelity PCR Master Mix 1000 x 20 ul reactions 480.00
MB119-B-S 2x Hot-Start High-Fidelity PCR Master Mix (Blue Dye) 100 x 20 ul reactions 80.00
MB119-B-L 2x Hot-Start High-Fidelity PCR Master Mix (Blue Dye) 1000 x 20 ul reactions 480.00
Description

MB119-B: 2x hot-start high-fidelity PCR Master Mix, with blue dye

The cheap 2x hot-start high-fidelity PCR Master Mix with non-interfering blue dye is ready-to-use 2x concentrated solution for DNA amplification and subsequent clonging/subcloning. Except DNA template and primers, the master mix contains all other components for hot-start high-fidelity PCR, including the hot-start high-fidelity DNA polymerase, dNTPs, MgCl2, reaction buffer, PCR stabilizer and enhancer at optimal concentrations for consistent and efficient PCR amplification. For a 20 ul PCR reaction, simply add 10 ul of 2x PCR Master Mix to specific primers, DNA template, and water. No additional loading buffer is required for electrophoresis when the PCR reaction is completed.

The 2x hot-start high-fidelity PCR master mix is based on the hot-start high-fidelity DNA polymerase. The hot-start high-fidelity DNA Polymerase is a new generation of high-fidelity DNA polymerase with improved ability of long fragment amplification, amplification specificity and yield. The hot-start high-fidelity DNA Polymerase can effectively amplify fragments up to 40 kb from simple templates such as λDNA and plasmids, up to 20 kb from complex templates such as genomic DNA, and up to 10 kb from cDNA templates. Its amplification mismatch rate is 1/53 that of ordinary Taq polymerase and 1/6 that of Pfu polymerase. With high resistance to PCR inhibitors, the hot-start high-fidelity DNA polymerase can be used for direct PCR of bacteria, fungi, plant and animal tissues, and even whole blood samples. The optimized hot-start high-fidelity DNA polymerase has rapid amplification speed (4 - 5 sec/kb) while maintaining high fidelity and yield. The optimized buffer system also helps the hot start high fidelity PCR master mix achieve high amplification specificity. The hot start high fidelity PCR master mix has excellent compatibility with crude samples, templates with uracil and GC-rich system (primer/template). The mix contains two types of monoclonal antibodies inhibiting the 5'→3' polymerase activity and 3'→5' exonuclease activity at room temperature, which enable it to perform hot start PCR with great specificity. Amplification will generate blunt-ended products, which are compatible with our homologous recombination-based one step DNA subcloning kit.

Optimal template and primer concentrations, and PCR program parameters should be determined experimentally by the investigator.

Features of the 2x hot-start high-fidelity PCR master mix with blue dye:
* Simple operation: The PCR master mix simplifies the assembly of PCR reaction and offers advantages of time savings, convenience, consistency, and minimal risk of contamination and pipetting errors. The tracking dye and precipitant have been added into the master mix so that the PCR product can be directly loaded for electrophoresis.
* Ultra-high fidelity: The fidelity is 53 times that of Taq DNA Polymerase and 6 times that of Pfu DNA Polymerase. For every 300,000 bases amplified, there are fewer than 5 mismatches.
* Fast amplification: normally 4-5 sec/kb; ~0.5 sec/kb if the amplification length is <1 kb; 4-150 times that of conventional PCR master mix.
* Long fragment amplification: up to 40 kb from simple templates such as λDNA and plasmids, up to 20 kb from complex templates such as genomic DNA, and up to 10 kb from cDNA templates.
* Wide adaptability: It is suitable for the amplification of various GC content fragments, has super tolerance to PCR inhibitors, and can be used for direct PCR of a variety of samples.
* Features of the hot-start high fidelity DNA polymerase (catalog No. MB118) contained in the 2x hot-start high-fidelity PCR master mix. 
* Low cost: $400 for 50 ml 2x hot-start high-fidelity PCR master mix. 

Applications of the 2x hot-start high-fidelity PCR master mix with blue dye:
* Routine PCR using genomic, viral and plasmid templates.
* RT-PCR.
* DNA cloning and subcloning.
* DNA mutagenesis.

Storage/Stability: Stable for 1 year at -20°C and 1 week at 4°C. For maximum recovery of the product, centrifuge the original vial after thawing and before removing the cap. Aliquot to avoid repeated freezing and thawing.

The hot-start high-fidelity DNA polymerase ($800 for 1000 units) contained in the 2x hot-start high-fidelity PCR master mixes is available alone.

Please contact us if you need protocol, datasheet, or SDS of our 2x hot-start high-fidelity PCR master mixes.

Quality of our 2x hot-start high-fidelity PCR master mixes has been proven by a number of publications citing our product.

PCR enzymes:
* Taq DNA polymerase.
* High yield Taq DNA polymerase.
* Hot-start high-fidelity DNA polymerase.
* Hot start Taq DNA polymerase with chemical or antibody inhibitor, used for SYBR green or Taqman probe-based real time PCR (qPCR or quantitative PCR).

2x PCR master mixes:
* 2x PCR PreMix with dye (green, red, blue).
* 2x rapid PCR Master Mix with a green dye.
* 2x long PCR Master Mix with a blue dye.
* 2x high yield PCR master mix with a blue dye.
* 2x hot-start high-fidelity PCR master mix with a blue dye.
* Blood direct PCR kit
* Cell and tissue direct PCR kit
* RT-PCR master mixes
* 2x qPCR master mixes
* RT-qPCR master mixes

PCR components and related products:
* dNTP mix, 10 mM each.
* dNTP mix, 25 mM each.
* Proteinase K powder.
* Proteinase K solution.
* Reverse Transcriptases.
* First strand cDNA synthesis kits.
* Murine RNase inhibitor.
* PCR enhancer.

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