Long PCR Taq polymerase Master Mix

Long PCR Taq polymerase Master Mix

DNK300P-01
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Master mix Long PCR Taq DNA Polymerase, the same high-performing DNA polymerases, now in the master mix format to provide you with convenience of use. The master mix is a special formulation containing the Long PCR Taq DNA polymerase. The Long PCR Taq was shown to amplify long templates from λ phage genome of up to 20 kb. It is also a better choice for amplifying complex targets, such as GC-rich targets.

Just add primers and template, your PCR reaction is ready to begin!

Long PCR Taq DNA Polymerase,a combination of two thermostable DNA polymerases, Taq and Pfu, is a special formulation designed for amplifying large fragment. This specially formulated Long PCR Taq was shown to amplify long templates from λ phage genome of up to 20 kb. It is also a better choice for amplifying complex template, such as GC-rich template.

Long PCR Taq is suitable as a direct replacement for ordinary Taq Polymerase in most applications. Using Long PCR Taq in your PCR reactions results in 3´-dA overhangs PCR products, which can be used in TA clone

Applications

• PCR amplification of DNA fragments any sizes around 5 kb

• DNA labeling

• DNA sequencing

• PCR for cloning

Features

• High fidelity: three times fidelity of Taq DNA Polymerase.

• Longer fragment: amplify long templates as long as 40kb.

• Amplification of complex template (GC rich or repetitive sequence).

• Generates 3'-dA and blunt end PCR products.

Note:

  • 10xLong PCR Bufferis classical Long PCR Taq DNA Polymerase buffer, is good for long template especially above 10kb.
  • 10xLong PCR Buffer is an alternatie long PCR buffer . It is for better fidelity but may not be robust for longer templates above 10kb.

Basic PCR Protocol

The following basic protocol serves as a general guideline and a starting point for any PCR amplification. Optimal reaction conditions

(incubation time and temperature, concentration of Taq DNA Polymerase, primers, Mg2+, and template DNA) vary and need to be optimized.

Notes on cycling conditions

- Initial denaturation can be performed over an interval of 1~5 min at 95 depending on the GC content of template.

-Denaturation for 30 sec to 2 min at 94~95 is sufficient. If the amplified DNA has a very high GC content, denaturation time may be increased up to 4 min.

-Optimal annealing temperature is 5 lower than the melting temperature of primer-temperature DNA duplex. If nonspecific PCR products are obtained optimization of annealing temperature can be performed by increasing temperature stepwise by 2.

-The number of PCR cycles depends on the amount of emplate DNA in the reaction mix and on the expected yield of the PCR products, 25-35 cycles are usually sufficient

for the majority PCR reaction. Low amounts of starting template may require 40 cycles.

-The time of the final extensi, on step can be extended for amplicons that will be cloned into T/A vectors.

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