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PCR

Maxime™ PCR PreMix (i-pfu)

Cat.No Capacity Inquire
25185 96 tubes Inquire
PRODUCT INFORMATION
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
Content 25185
Maxime™ PCR PreMix Kit(i-pfu, for 20 μl rxn) 96 tubes

 

 

Technical Data

Amplification test results with i-pfu DNA Polymerase

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

Sensitivity test

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

TroubleShooting Guide
QWhen I look at the manual for the experiment, only the experiment is shown when the reaction volume is 50 μl. I want to change the volume to 20μ Can I put it?
A If the PCR reaction volume is reduced from 50 μl to 20 μl, the same amount of Taq should be added. You can experiment with the same amount of template and primer. Do not add 5μl of 10X PCR buffer and reduce to 2μl.
QI am currently performing full genomic sequencing.
AOur i-pfu has an error rate of 1X10-6 and can amplify up to 10kb.If you want a product with higher functionality, we recommend the i-StarMAXTM II product, which has an error rate of 1.8X10-8 and can synthesize up to 20kb. 
QHIV mutagenesis is under study. point mutation, but the kit is expensive and I want to buy only the enzyme. Is there a suitable product?
A i-pfu™ DNA pol.ymerase is recommended.i-pfu DNA polymerase has a proof-reading activity that lowers the rate of mutation during the PCR reaction. 
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