RANGER Mix

Description

A unique blend of high-performance hot-start MyTaq™ HS DNA Polymerase and a proprietary proofreading enzyme, specifically developed for long PCR applications

Product Highlights

Efficient – specifically developed to give reliable amplification of fragments between 10 kb and 25 kb in length
Specific – incorporates an antibody-mediated hot-start enzyme blend that remains completely inactive during PCR set-up to prevent non-specific amplification
Fast – rapid enzyme activation and highly efficient amplification support reduced time to results
Flexible – ideal for amplifying a broad range of large fragments from even complex targets including human genomic DNA
Accurate – possesses 3’ – 5’ proofreading exonuclease activity for increased fidelity, thereby enabling cloning of long PCR products

Product Description

RANGER Mix is an easy-to-use, high-performance enzyme blend, specifically developed to amplify fragments up to 25 kb in length. RANGER contains a unique combination of a highly-efficient DNA polymerases and novel buffer system that deliver the improved efficiency necessary for reliable amplification of longer amplicons.

RANGER is an antibody-mediated hot-start enzyme blend that eliminates non-specific amplification during reaction set-up. The inactivated enzymes do not possess polymerase activity, thereby preventing the non-specific amplification, such as primer-dimer formation, that often hinder long PCR reactions from the start.

RANGER Mix is an all-in-one mastermix containing dNTPs, MgCl₂ and enhancers at optimal concentrations, minimizing the requirement for PCR optimization and driving greater sensitivity. RANGER Mix also improves reproducibility and minimizes the requirement for assay repeats, by reducing the number of pipetting steps, and therefore the risk of manual error, during reaction set-up.

Applications

Long PCR
TA cloning

Improved yield and specificity in long PCR.

PCR Enzyme Guide

Download the PCR Enzyme Guide with detailed product descriptions and performance data to help you choose the best product for your research

PCR Selection Chart

Select the best reagent for your research

Having struggled with PFU and other polymerases, I was extremely happy to try RANGER DNA polymerase and it worked for both 8.5 kb and 6.5 kb.

NWCR Institute, Bangor University, UK

Product Selection

Please refer to the PCR Selection Chart to confirm the recommended product for your PCR application.

Resources

Documents

RANGER Mix - Fact Sheet RANGER Mix - MSDS RANGER Mix - Product Manual

PCR Enzyme Guide PCR Selection Chart

Certification of Analysis (COAs)

BIO-25052_PM361-B077840 BIO-25052_PM361-B074140 BIO-25052_PM361-B074070 BIO-25052_PM361-B067310 BIO-25052_PM361-BO65090

ViewHide all 33 COAs

Specification

Components

Reagent	500 Reactions
RANGER Mix, 2x	10 x 1.25 mL

Volume

BIO-25052: 500 x 50 µL Reactions: 10 x 1.25 mL

Concentration

2x

Storage & Stability

All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided. When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date indicated on the outer box label.

Components may also be stored at +4°C if required, although storage at -20°C is recommended. When stored at +4°C, reagents will remain stable for a period of 2 weeks from date of receipt.

Shipping conditions

Shipped on Dry Ice or Blue Ice

FAQs

What is the final working concentration (at 1x) of MgCl₂ in each mix?

Bioline's mixes contain magnesium chloride at the following concentrations (please see table below). An additional tube of 50 mM MgCl₂ solution is supplied with each mix to facilitate further optimization if required.


Bioline Mix	Final Magnesium Concentration

ACCUZYME Mix	2.0 mM.

BioMix / BioMix Red	2.5 mM.

ImmoMix / ImmoMix Red	3.0 mM.

BIO-X-ACT Short Mix	2.0 mM.

MangoMix	2.5 mM.

MyTaq	3.0 mM.

RANGER	1.5 mM.

Which polymerase do I need for my specific application?

At Bioline we pride ourselves in supplying high-quality polymerases to suit your requirements. To aid your selection of the most suited enzyme for your specific applications, please see our enzyme selection tool.

What are the storage conditions and stabilities of Bioline polymerases?

All our polymerases are guaranteed for a period of 12 months from the date of purchase. These should be stored at -20°C during this time for optimal retention of activity.

Please Note: We do not recommend the storage of our polymerases at -80 °C as ice crystals could form on the active site, which may affect or destroy the activity of the enzyme.

I am having problems optimizing my PCR, what would you recommend?

PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.

Observation	Recommended Solution(s)
No or low PCR yield	Enzyme concentration too low – increase the amount of enzyme in 0.5 U increments.
	Primers degraded – check quality and age of the primers.
	Magnesium concentration too low – increase concentration in 0.25 mM increments with a starting concentration of 1.75 mM.
	Primer concentration not optimized. Titrate primer concentration (0.3-1 µM); ensuring that both primers have the same concentration.
	Template concentration too low – Increase concentration of template.
	Perform a positive control to ensure that the enzyme, dNTPs and buffers are not degraded and/or contaminated.
Multiple Bands	Primer annealing temperature too low. Increase annealing temperature. Primer annealing should be at least 5°C below the calculated Tm of primers.
	Prepare master mixes on ice or use a heat-activated polymerase.
	For problems with low specificity. Try adding 3% DMSO (not supplied) to improve specificity.
Smearing or artifacts	Template concentration too high. Prepare serial dilutions of template.
	Too many cycles. Reduce the cycle number by 3-5 to remove non-specific bands.
	Enzyme concentration too high - decrease the amount of enzyme in 0.5 U increments.
	Extension time too long. Reduce extension time in 0.5-1 minute increments.

What do the terms yield, fidelity, processivity and specificity actually mean?

These terms refer to parameters to be considered when performing PCR and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial: Yield: The amount of DNA produced in a PCR reaction. Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand. Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified. Specificity: A measure of the unwanted by-products generated in a reaction.

What are the benefits of using a hot-start polymerase?

During PCR setup at room temperature, standard polymerases will have some activity. When using a hot-start polymerase, this enzyme will have no activity at room temperature, thus reducing the risk of unspecific products in the reaction due to these mis-primed oligonucleotides. This feature makes this type of polymerase ideally suited to high-throughput applications, where the reactions may be left sitting at room temperature for prolonged periods of time.

What is a proofreading polymerase?

In nature, the ability of a DNA polymerase to correct misincorporations of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed "proofreading activity" and occurs in the 3' to 5' direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3'end (A-overhangs), creating blunt ends.

How is one unit of activity of a polymerase defined?

One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C.

What are the advantages of using a 2x mix rather than setting up a PCR reaction from scratch?

All Bioline polymerases are available in the convenient format of a 2x master mix, containing all the reagents and additives necessary to perform successful PCR, with the exception of template, primers and water. This formulation not only provides a convenient and hassle-free PCR setup, but also significantly reduces the chances of human error, inaccuracies and contamination by reducing the pipetting steps required.

What is the recommended reaction volume of the polymerase mixes?

We recommend a final reaction volume of 50 µL, this requiring the use of 25 µL of the 2x master mix and to make up to 50 µL using template, primers and PCR grade water.

What is the composition of the polymerase mixes?

Whilst the exact composition of the mixes is proprietary information, all mixes are made up of reaction buffer, magnesium, dNTPs, polymerase and additives, designed to keep the polymerase stable in the mix, as well as provide the optimal conditions for PCR.

What is the concentration of the polymerase mixes?

All Bioline PCR mixes come supplied at a 2x concentration, requiring the user simply to add template, primers and PCR grade water to a final concentration of 1x.

My DNA sample contains PCR inhibitors, which impairs my PCR results. What can I do for optimization?

If possible, we would recommend an additional cleanup of affected samples, for instance, customers had very good experiences with the clean-up of samples containing humic acids with SureClean Plus. Alternatively decreasing the template concentration will help to minimize the amount of inhibitors. The amount of primer can be increased, but do not exceed the suggested limits.

Is a sample available?

Please click here in order to request your sample. You will receive an email confirmation within two business days with delivery details.

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