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Some applications of this product  may require a license which is not provided by the purchase of this product.

For research use only. 

Ready to load PCR Master Mix for routine applications

  • suitable for templates up to 5 kb
  • reaction set-up and shipment without dry ice
  • all-in-one master mix format reduces pipetting errors and saves time

Ordering

MgCl2 (mM) final concentration
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Catalog number

04-12-00S25

Quantity

€ 0.00

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Details

Overview

Description

Optimized ready-to-use PCR master mix for routine PCR assays. This master mix contains thermostable Taq polymerase FIREPol®, MgCl2, dNTPs and buffer with detergent. You just need to add template, primers and water.

Ready to Load format also includes a compound needed for direct loading onto agarose gel and two tracking dyes (blue and yellow) that allow to monitor progress during electrophoresis.

We recommend using 5x FIREPol® Master Mix Ready to Load in any PCR application that will be visualized by agarose gel electrophoresis and ethidium bromide staining. 5x FIREPol® Master Mix Ready to Load is not recommended for use in post-PCR applications where spectro-photometric measurements (absorbance or fluorescence) are necessary because yellow and blue dyes can interfere with these applications.

Properties

Concentration: 5x

Hot-start: no

Ready to load: yes

Fidelity: same as Taq

Cloning Type: T/A Cloning

Amplicon Size: up to 5 kb

Mix Components

FIREPol® DNA polymerase: highly processive. thermostable DNA polymerase

5x Reaction Buffer B0.4 M Tris-HCl, 0.1 M (NH4)2SO4, 0.1% w/v Tween-20

12.5 mM MgCl21x PCR solution – 2.5 mM MgCl2

1 mM dNTPs of each: 1x PCR solution – 200 µM dATP, 200 µM dCTP, 200 µM dGTP and 200 µM dTTP

Blue dye: migration equivalent to 3.5-4.5 kb DNA fragment

Yellow dye: migration equivalent to 35-45 bp DNA fragment

Compound that increases sample density for direct loading

Plant genomic DNA
Related Resources

FAQ

Which polymerases / master mixes are compatible with a downstream restriction enzyme digest without cleaning up the PCR reaction?

FIREPol® and HOT FIREPol® DNA Polymerases and Master Mixes, as well as SolisFAST® Master Mix are compatible with most restriction enzymes without cleaning up the PCR reaction product.

SolisFAST® Master Mix with UNG contains dUTP instead of dTTP leading to the generation of PCR products containing uracil. Check your restriction enzyme for the ability to digest U-containing DNA amplicons.

Which DNA polymerases / master mixes should I use to obtain PCR product suitable for Sanger sequencing?

Products generated by our FIREPol®, HOT FIREPol® DNA Polymerases and Master Mixes, as well as SolisFast® Master Mixes may be used for Sanger sequencing.

Prior to the sequencing, the products should be purified to remove excess primers and nucleotides (enzymatically with ExoI/SAP or Spin column-based nucleic acid purification).

Products generated with 5x HOT FIREPol® Blend Master Mix should be cleaned up utilising column-based nucleic acid purification method, because this mix contains proofreading enzyme with 3’-5’ exonuclease activity and may degrade your sequencing primer.

Please note that Ready to Load versions of our Master Mixes are not recommended for use in applications where spectro-photometric measurements (absorbance or fluorescence) are necessary because tracking dyes can interfere with these applications.

Which DNA polymerases / master mixes should I use to obtain PCR product suitable for Sanger sequencing?

Products generated by our FIREPol®, HOT FIREPol® DNA Polymerases and Master Mixes, as well as SolisFast® Master Mixes may be used for Sanger sequencing.

Prior to the sequencing, the products should be purified to remove excess primers and nucleotides (enzymatically with ExoI/SAP or Spin column-based nucleic acid purification).

Products generated with 5x HOT FIREPol® Blend Master Mix should be cleaned up utilising column-based nucleic acid purification method, because this mix contains proofreading enzyme with 3’-5’ exonuclease activity and may degrade your sequencing primer.

Please note that Ready to Load versions of our Master Mixes are not recommended for use in applications where spectro-photometric measurements (absorbance or fluorescence) are necessary because tracking dyes can interfere with these applications.

What is the function of MgCl₂ in a PCR reaction?

The concentration of the MgCl2 in PCR reaction affects the productivity of your PCR reaction. Higher concentrations of MgCl2 may increase productivity of DNA polymerase, but also may decrease specificity causing unexpected products.

Usually, 1.5-2.5 mM final concentration of MgCl2 is optimal for general PCR. However, optimal concentration of MgCl2 depends on template, buffer, DNA and dNTPs. Solis BioDyne produces Master Mixes at different MgCl2 concentration, which allows greater flexibility in reaction optimisation. For your very first experiment with Master Mix, we recommend to use Master Mix with 1,5 - 2 mM MgCl2 (in final 1x PCR Mix)

What are the storage conditions and stability of Solis BioDyne reagents?

Solis BioDyne products should be stored at -20°C.

Shipping and temporary storage for up to 1 month at room temperature (*1525°C) has no detrimental effects on the quality of Solis BioDyne reagents.

Freeze-thaw stability is tested for each product. Most PCR and qPCR products have passed 30 freeze-thaw cycles with no changes in performance. Specific information is found in Storage and Shipping conditions of each product.

When stored and handled under the recommended conditions, full activity of the reagents is retained until the Expiry Date printed on the tube label.

*World Health Organization (2003). Guidelines for the Storage of Essential Medicines and Other Health Commodities.

Storage & Shipping
Storage

Routine storage: -20ºC

Temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of FIREPol® Master Mix Ready to Load

Shipping

At room temperature