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IMMOLASE™ DNA Polymerase

Ordering

Cat. No.
Size
BIO-21046
250 Units
BIO-21047
500 Units

*link will take you to our exclusive distribution partner site

Description

IMMOLASE™ is a heat-activated, thermostable DNA polymerase. IMMOLASE provides high yield and improved specificity when compared to standard polymerases and can eliminate the presence of non-specific binding, such as primer-dimers and mis-primed products.

Product Highlights

  • Higher Performance - robust polymerase
  • Chemical hot-start PCR enzyme - eliminates non-specific binding
  • Reaction requires activation step - allows convenient reaction set-up at room temperature
  • Suitable for TA cloning - leaves ´A´ overhang

Product Description

IMMOLASE™ is a heat-activated thermostable DNA polymerase and provides high yield and improved specificity when compared to standard polymerases. IMMOLASE can eliminate the presence of non-specifics such as primer-dimers and mis-primed products.

IMMOLASE is inactive at room temperature and requires activation by heat treatment for 10 minutes prior to PCR cycling. This provides greater flexibility in reaction set ups and allows the premixing of PCR reagents at room temperature. Subsequently, the reaction can be handled according to preferred protocols for thermostable DNA polymerases.

Applications

  • For multiplex reactions
  • Products suitable for TA cloning
Main

PCR Enzyme Guide

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PCR Selection Chart

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Specification

Components

Product

250 Units

500 Units

IMMOLASE DNA Polymerase

1 x 50 µL

1 x 100 µL

10x ImmoBuffer

 1.2 mL

2 x 1.2 mL

50 mM MgCl2 Solution

 1.2 mL

1 x 1.2 mL

Concentration

5 u/µL

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.

Shipping conditions

Shipped on Dry Ice or Blue Ice.



Resources

Documents

IMMOLASE DNA Polymerase - Manual
IMMOLASE DNA Polymerase - MSDS
PCR Enzyme Guide - A New Generation of End-Point PCR
IMMOLASE DNA Polymerase - Fact Sheet

Latest Certification of Analysis (COAs)

BIO-21048_PL351-B129840

FAQs

What is the heat activation time required for this polymerase?

IMMOLASE DNA Polymerase requires a heat-activation step of 10 minutes at 95°C.



Which specific applications is IMMOLASE DNA Polymerase suited to?

As well as most standard applications, IMMOLASE DNA Polymerase is ideally suited to the following:

-          High-throughput applications

-          Multiplex PCR

-          TA Cloning

IMMOLASE DNA Polymerase has been manufactured under 13485 Quality Management System and is suitable for further manufacturing use as an IVD component.



What are the features of IMMOLASE DNA Polymerase?

The features of IMMOLASE DNA Polymerase are as follows:

  • Heat Activation: 10 minutes at 95°C
  • Speed: 15-30 s/kb (template dependant)
  • Length amplified: Around 5 kb (genomic DNA)

Optimal Extension Temperature: 72°C



Which polymerase do I need for my specific application?

At Meridian 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 Meridian 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.

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 Meridian 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 would be the recommended DNA polymerase for difficult samples like GC rich or bisulfite converted DNA?

The MyTaq HS DNA polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. Especially the hot-start is important, as it decreases primer dimer and unspecific amplification. We would suggest to start with the recommended protocol. If optimization is needed, we would suggest to optimize the annealing temperature. Due to the degraded DNA after bisulfite conversion it may be useful to increase the amount of template and polymerase.

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.

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