ELISA Long PCR Taq polymerase Master Mix

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 formµLation 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 formµLation designed for amplifying large fragment. This specially formµLated 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 resµLts in 3´-dA overhangs PCR products, which can be used in TA clone Applications • PCR amplification of DNA fragments any Quantitys 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 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.