2X PCR Master mix Solution (i-pfu)
2x PCR Master mix type product with high purity pfu DNA Polymerase with low error rate and Proofreading function which is essential for gene cloning requiring high accuracy.
– All reagents required for the reaction are in solution in the tube
– Immediate PCR by adding template and primer
• PCR reaction volume control
• High accuracy
– Proof-reading activity enables amplification of long DNA template
– Optimal for experiments requiring accuracy such as gene cloning and expression
• Low error rate
– 3’→5’ Exonuclease is a typical thermostable proof-reading enzyme with an error rate of 1×10-6
• Blunt-end cloning possible
2x PCR Master Mix Solution (i-pfu) is a product designed to achieve the best and simple result by using 2x Master mix type in one tube for all components necessary for PCR reaction including i-pfu DNA Polymerase. This product is able to perform PCR by adding template DNA, primer set and D.W only, and gel loading buffer is included, so gel loading can be done without any other treatment. The i-pfu DNA Polymerase used in this product is a typical heat-resistant proof-reading enzyme with 3 ‘→ 5’ exonuclease activity. It has high fidelity and has an error rate of 1×10-6 (one error rate per 13,000 base pairs) . The proof-reading activity of pfu DNA Polymerase is that when a mismatched nucleotide interferes with DNA polymerase during polymerization, it can be removed and corrected with a complementary nucleotide. This activity allows PCR products of more accurate base sequences to be obtained than Taq DNA Polymerase, but allows amplification of longer DNA templates compared to Taq DNA Polymerase. This is because the mismatched nucleotide interferes with the polymerization reaction. The Pfu DNA Polymerase can remove the DNA polymerase from the template during the DNA synthesis process. It is suitable for long-size PCR (up to ~ 30 kb) which is difficult to amplify. It has high fidelity and proof reading function compared with general Taq DNA Polymerase. As a result, DNA fragments longer than Taq DNA Polymerase (up to 30 kb) can be amplified. Because of these properties, Pfu DNA Polymerase is widely used in experiments where accuracy is required, such as gene cloning and expression, although polymerization efficiency is low and it takes more time to polymerize. The 2x PCR Master Mix Solution (i-pfu) has high fidelity using the i-pfu DNA Polymerase and features a low error rate of 1×10-6 (one error rate per 13,000 base pairs). In addition, it is highly reproducible and can be tested quickly and easily for various samples.
Sensitivity based on various size
2x PCR Master Mix Solution (i-pfu) and products of the same function of other companies were used to confirm the PCR efficiency by template concentration for various kinds of DNA by electrophoresis. We amplified 4.5 Kb of E. coli-derived DNA as a template and amplified 1 Kb fragment of lambda DNA as a template.
A, 2x PCR Master mix Solution(i-pfu); B, Supplier A; C, Supplier B
Lane M, 100 bp DNA Marker, 1 Kb DNA Marker; lane N, Negative control