Products

Isotherm2G DNA polymerase

(currently out of stock until further notice, please request)

 

is a mesophilic DNA polymerase, which synthesizes DNA from both DNA and RNA templates with a high strand displacement activity. Isotherm2G is the next generation of the original Isotherm DNA polymerase with further improved reverse transcription activity. The temperature optimum (between 55-70°C) renders Isotherm2G DNA polymerase the choice No. 1 for isothermal amplification reactions such as loop-mediated isothermal amplification (LAMP), strand displacement amplification (SDA), rolling circle amplification (RCA), ramification amplification (RAM), genome amplification and more.

 

Article Product Size Price
#8801S Isotherm2G┬áDNA polymerase 1600┬áU, 8U/┬Ál, 200 ┬Ál ÔéČ 75,-
#8801M Isotherm2G DNA┬ápolymerase 8000 U, 8U/┬Ál, 1000┬á┬Ál ÔéČ 340,-

Bulk offer: Should you require larger amounts (> 10,000 U), significant discounts are available. This product can also be customized. Please contact us to receive an individual quotation.

 

download-pdf Product Manual (PDF)Order

 

 

Example Results

Isothermal amplifications  

It's that simple: just an incubation at 59°C for 30min resulted in the amplification of both a DNA target (red curve) and a RNA target (blue curve), using the LAMP method. Isotherm can be used for RCA, SDA, RAM, LAMP and many other amplification technologies.

 

 

 

 

Isotherm DNA polymerase also has a very good reverse transcription activity. After amplification, a melting curve measurement can be performed to confirm the specific product generation. DNA and RNA detections can thus be undertaken allowing an easy reaction setup, a fast reaction time and a close-tube system.

 

 

 

Applications

Applications

  • Rapid, isothermal detection and identification of DNA and RNA targets
  • Reverse transcription
  • Real-time isothermal detections
  • LAMP, SDA, RCA, RAMP
  • Primer extensions

Reaction Setup

Reaction setup

Component Volume Final concentration
Primer mix (various concentrations) various ┬Ál various
dNTPs (10 mM) 1┬á┬Ál 400 ┬ÁM
10 x┬áIsotherm2G reaction buffer 2.5 ┬Ál 1x
Isotherm DNA polymerase┬á8 U/┬Ál 1 ┬Ál 8 U / reaction
Template/Sample extract various┬á┬Ál ┬ávarious
Nuclease-free water up to 25 ┬Ál total reaction volume ┬á

Keep all components on ice.
Spin down and mix all solutions carefully before use. 

Primer mixes and concentration depends strongly on the employed isothermal reaction method.

Isotherm2G and Isotherm2G reaction buffer does not include a fluorescent dye. For for real-time measurements please add an appropriate amount of a suitable dye.

 

 

Detailed product description

 

Content

Isotherm DNA polymerase is supplied as a 8 U/┬Ál solution containing glycerol. It comes together with a 10x reaction buffer optimized for isothermal amplifications.

 

Quality Control

DNA polymerase activity: Isotherm activity has been monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and DNA primer.
Enzyme concentration has been determined by protein-specific staining. Please inquire more information at info@mypols.de for the lot-specific concentration.

 

Legal information

Certain isothermal amplification methods may be patent protected. The purchase of this product does not convey to the purchaser a legal right to use the purchased product to perform any isothermal amplification methods for any commercial purposes.

 

Storage

This product is shipped on cool packs. Please store the product upon arrival at -20┬░C.

 

 

img-faqsFAQ`s

How can I optimize the assay conditions?

Most variable paramters are the incubation temperature, dNTP and Mg2+ concentration.

How much Mg2+ is in the Isotherm2G reaction buffer?

Final 1x reaction buffer contains 3.25 mM magnesium ions.

Which temperature shall I use?

 

The optimal reaction temperature strongly depends on the primer sequences and the amplification method. To start with, we would recommend to try a temperature gradient in order to find the best reaction temperature for your assay. Isotherm2G is most active between 55-70°C. Isotherm2G can be heat-inactivated upon incubation at temperatures above 70°C. 


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