Description

Dried & aliquoted PreMix Kit with high purity pfu DNA Polymerase with low error rate and Proofreading function which is essential for gene cloning requiring high accuracy 

• • Dried & aliquoted PreMix, All-in-one products
     - All the reagents required for the reaction are in dried pellet in one tube   
     - Immediate PCR by adding template and primer
     - Gel loading electrophoresis is possible because it contains loading dye
• • Excellent product stability and reproducibility 
     - Excellent reproducibility with dried & aliquoted PreMix by ALHP system  
     - Excellent stability due to oxidation and moisture prevention by vacuum compression manufacturing method 
• • 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 1x10^-6
• • Blunt-end Cloning possible

Maxime™ PCR PreMix Kit (i-pfu) contains i-pfu DNA Polymerase and all the components necessary for PCR reaction (dNTP Mixture, reaction buffer, etc.) are included in each PCR tube by ALHP system. It is very excellent in reproducibility because it can reduce the experimental error to pipetting between the experimenters. It is a product with excellent stability of activity due to oxidation and moisture prevention of major components in the product by vacuum compression manufacturing method. In addition, this product can perform PCR by adding template DNA, primer set and D.W only, and it can contain gel loading buffer.

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 a low error of 1x10^-6 (one error rate per 13,000 base pairs) rate. 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 Maxime™ PCR PreMix Kit (i-pfu) has high fidelity, low error rate, excellent stability and reproducibility and has all the components needed for PCR reactions using i-pfu DNA polymerase. It is a dry pellet in the tube, so it can be tested very quickly and easily for various samples.

Applications

• 01General PCR
• 02template amplification for sequence analysis
• 03Blunt Cloning

Kit Contents

Technical Data

As a result, it was confirmed that the i-pfu DNA polymerase and 1 Kb (lamda DNA template) and 4.5 Kb (5F' plasmid DNA) amplification were used to confirm the sensitivity of Maxime ™ PCR PreMix Kit (i-pfu) pfu DNA Polymerase.

[ 1 Kb ] Lane M, 1 Kb DNA Marker; lane 1, 200 pg DNA; lane 2, 20 pg DNA, lane 3, 2 pg DNA
[ 4.5 Kb ] Lane M, 1 Kb DNA Marker; lane 1, 50 ng DNA; lane 2, 10 ng DNA, lane 3, 2 ng DNA

MaximeTM PCR PreMix Kit (i-pfu) was tested for the sensitivity test. As a result of checking by 4.5 Kb (5F' plasmid DNA) and 9 Kb (SLT plasmid DNA) amplification, both sizes are superior to those of other products.

[ 4.5 Kb ] Lane M, 1 Kb DNA Marker; lane 1, 200 ng DNA; lane 2, 100 ng DNA, lane 3, 50 ng DNA; lane 4, 25 ng DNA; lane 5, 12.5 ng DNA, lane 6, 6.25 ng DNA
[ 9 Kb ] Lane M, 1 Kb DNA Marker; lane 1, 250 ng DNA; lane 2, 50 ng DNA, lane 3, 10 ng DNA; lane 4, 2 ng DNA; lane 5, 400 pg DNA