BLIRT LongNova-RED DNA Polymerase

LongNova polymerase with red dye for direct gel loading

LongNova-RED DNA Polymerase
Name Part Number Pack Size
LongNova-RED DNA Polymerase RP281R 200 U
  RP282R 1000 U

 

LongNova-RED DNA Polymerase is a mixture of thermostable Pwo and Taq polymerases with the addition of an inert red dye which facilitates accurate low volume pipetting and is an indicator of enzyme addition. The dye has no adverse effect on the outcome of PCR; yields are the same as with the standard LongNova DNA Polymerase.

The thermostable LongNova DNA Polymerase catalyses DNA synthesis in a 5’-3’ direction and shows a 3’-5’ exonuclease activity. It is characterized by high processivity and proofreading properties. It is ideal for long range PCR (up to 20 kb) and GC-rich template amplification applications.

Use of the LongNova-RED DNA Polymerase decreases the risk of error during a reaction set up, e.g. skipping the polymerase or inaccurate reagent mixing. In addition, the PCR product can be applied directly to a gel after amplification without the need to mix it with a loading buffer. This reduces the time required to perform agarose or polyacrylamide electrophoresis.

Features and advantages

- Wide range of product sizes – from 2 to 20 kb

- High proofreading properties (3'-5’ exonuclease activity)

- Facilitate PCR reaction set-up

- Direct gel loading

Applications

- Long PCR products

- Molecular cloning

- Site-directed mutagenesis and other methods which require high fidelity

Storage buffer

20 mM Tris-HCl (pH 7.4, 25°C), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 2.5 mg/ml BSA, inert dye, 50% glycerol

Quality control

Free of unspecific nucleases and DNA contamination. Extensively tested in various long range PCR reactions.

Unit definition

One unit is defined as an amount of enzyme required to incorporate 10 nmol of dNTPs to an insoluble DNA fraction in 30 minutes at 75°C in a 50 μl reaction.

Additional information

- The recommended concentration of LongNova-RED DNA Polymerase in the reaction is 0.05 U/µl. If other polymerase concentrations are desired, magnesium ion concentration optimisation may be required. The table below presents the Mg2+ concentration calculations in line with the volume of 50 mM MgCl2 added to the reaction.

- The minimal volume of the polymerase taken into a reaction is 1.5 µl / 50 µl, which relates to efficient sample loading to a gel.

- PCR products may be applied directly to a gel in a volume range of 5 - 15 µl with no need to mix them with a DNA loading dye as well. Electrophoresis, detection and visualization may be carried out in line with standard procedures.

Storage conditions

Store all components at -20°C.

Shipping conditions

Shipping on dry ice

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