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
• PCR amplification of DNA fragments any sizes around 5 kb
• DNA labeling
• DNA sequencing
• PCR for cloning
• 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.
- 10xLong PCR BufferⅠis classical Long PCR Taq DNA Polymerase buffer, is good for long template especially above 10kb.
- 10xLong PCR Buffer Ⅱ 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.