The Thermo Scientific Extensor Long Enzyme Mix provides superior yield and sensitivity for long fragment PCR applications. Standard Taq DNA polymerase incorporates 1 error in every 9kb. When amplifying fragments that are close to 9kb or larger, it is important to ensure that you are amplifying with no errors. The Extensor Long PCR Enzyme Mix is able to amplify fragments up to and greater than 20kb with 6 times the fidelity and double the yield of Taq.
Product Detail
Applications
Long range PCR amplification of fragments >9kb
High fidelity PCR
Amplification of difficult and GC rich fragments
Cloning
Mutagenesis
Description
Extensor Long PCR Enzyme Mix is a blend of ThermoPrime Taq DNA Polymerase with a proprietary thermostable proofreading enzyme. The two enzymes act synergistically to generate larger products than either enzyme individually (Up to 20kb). This makes the enzyme blend ideal for the full-length amplification of long sequences for cloning. The majority of the activity in the Extensor Long PCR Enzyme Mix comes from ThermoPrime Taq DNA Polymerase. Despite the 3’ to 5’ exonuclease (proofreading) activity present in the mix, sufficient PCR products will be generated with an A-overhang for T/A cloning (Mung bean nuclease can be used on products to remove the A-overhang for blunt end cloning).
Two buffer systems are available for application specific optimization:
Extensor Buffer 1 is used for amplifying fragments up to 12kb.
Extensor Buffer 2 is used for amplicons longer than 12kb or problematic PCR.
The enzyme is available in an easy to use master mix format; the Extensor PCR Enzyme Mix, reaction buffer, dNTPs and MgCl2 are all present in the mix. Master mixes are also available in a ReddyMix format, which includes an inert red dye and precipitant for direct gel loading of the PCR product. These convenient master mix formats reduce the risk of contamination and reagent handling errors.
Features
Synergistic enzyme mix to amplify DNA templates up to 20kb
6 times higher fidelity than standard Taq DNA polymerase
Double the yields of standard Taq DNA polymerase
Two buffer systems for application specific optimization
Proofreading enzyme has 5' to 3' exonuclease activity
Storage at +4°C eliminates freeze/thawing of individual components
Available in master mix format and ReddyMix format for significant time saving during PCR setup
Kit Contents for Core Kits
Vial of Extensor Enzyme Blend (5U/μl)
Vial of Extensor 10X Buffer 1*
Vial of Extensor 10X Buffer 2*
Vial of 25mM MgCl2
* The 10X buffers both contain 22.5mM MgCl2 and dNTPs. An additional vial of 25mM MgCl2 is provided for any optimization that may be necessary.
Also available with Lambda DNA and primers to be used as a control.
Kit Contents for Master Mixes
Vial of Extensor Long PCR Master Mix (4 versions available - see table below).