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PCR

2X PCR Master mix Solution (i-MAXⅡ)

Cat.No Capacity Inquire
25266 0.5 ㎖ x 2 vials Inquire
PRODUCT INFORMATION
Description

2x PCR Master mix type with high sensitivity, accuracy and low error rate, capable of amplification from small size DNA to DNA of more than 20 Kb with a proofreading function   

  • • Ready to Use
       - 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 and low error rate   
       - Proof-reading activity enables amplification of long DNA template    
       - Ensures high amplification efficiency for amplification of long and short template DNA
  • • Cloned DNA, human genomic DNA, as well as GC-rich or looped sequences that are difficult to amplify  AND applicable to various DNA  

2x PCR Master Mix Solution (i-MAXTM II) is a product designed to achieve the best results with simple 2x master mix type in one tube, including all components necessary for PCR reaction including i-MAXTM II DNA Polymerase is. 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-MAXTM II DNA Polymerase used in this product has the advantages of Taq DNA Polymerase and Pfu DNA Polymerase to overcome the disadvantages of each enzyme and to obtain new functions. It has a proofreading function, Kb) to 20 Kb or more.

The advantages and disadvantages of each enzyme are as follows. In the case of Taq DNA Polymerase, if the template is over 5 Kb, the efficiency of amplification is significantly lowered. Depending on the type of template, DNA template of 10 Kb or more is difficult to amplify. This is because Taq DNA Polymerase has no proof-reading activity, and when mismatched nucleotides are inserted during DNA synthesis, the template fails to polymerize. These defects in Taq DNA Polymerase can be overcome by using a thermostable DNA polymerase with corrective function. This corrective function is to remove the mismatched (non-complementary) nucleotide when the DNA Polymerase undergoes a polymerization reaction, and correct it 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.

Pfu DNA Polymerase is a typical enzyme of thermostable DNA Polymerase with this corrective function. It is suitable for long size PCR (up to ~ 30 Kb), which is difficult to amplify with general Taq. It has higher fidelity and proof reading function than general Taq DNA Polymerase . However, Pfu DNA Polymerase has the disadvantage that it takes more time to polymerize the DNA due to its poor polymerization efficiency. As mentioned above, the 2x PCR Master Mix Solution (i-MAXTM II) uses i-MAX ™ II DNA Polymerase, which takes advantages of Taq DNA Polymerase and Pfu DNA Polymerase, It is a product that can increase the amplification rate in size, show excellent results in sensitivity part, reproducibility is high, and can be tested quickly and easily on various samples.

Applications
  • 01General PCR
  • 02Human genomic DNA for PCR
  • 03Long DNA fragment amplify to 20kb
  • 04TA Cloning & Blunt Cloning, etc.
Kit Contents
Content 25266
2x PCR Master mix Solution(i-MAXTM II) 0.5 ml x 2 tubes

 

 

Technical Data

Sensitivity comparison data

In order to compare the sensitivity of 2x PCR Master Mix Solution (i-MAX TM II) with other products of the same function, 1 Kb DNA fragment PCR was performed by diluting human genomic DNA template with concentration. It shows that our kit obtained better result.

A, Competitor; B, 2x PCR Master mix Solution(i-MAXTM II )
Lane M, 1 Kb DNA Marker; lane NC, Negative control; lane 1, 50 ng gDNA, Lane 2, 25 ng gDNA; lane 3, 12.5 ng gDNA; lane 4, 6.25 ng gDNA; lane 5, 3.125 ng gDNA; lane 6, 1.56 ng gDNA; lane 7, 0.753 ng gDNA

In order to compare the sensitivity of 2x PCR Master Mix Solution (i-MAXTM II) with other products of the same function, long PCR of 20 Kb fragment was performed by diluting λDNA template by concentration. It shows that our kit obtained better result.

A, 2x PCR Master mix Solution(i-MAXTM II); B, Competitor
Lane M, 1 Kb DNA Marker; lane N, Negative control; lane 1, 50 ng DNA; lane 2, 25 ng DNA, lane 3, 12.5 ng DNA, lane 4, 6.25 ng DNA, lane 5, 3.125 ng DNA; lane 6, 1.56 ng DNA; lane 7, 0.753 ng DNA

TroubleShooting Guide
Q i-MAXTMII used a PCR pattern similar to the previous one while searching for taq for TA cloning by extracting genomic DNA from the plant, but the cloning does not work well.
AOur i-MAXTMII is designed for long PCR, but TA cloning is also available. However, when TA cloning small size products, we recommend i-Taq™, which is suitable for TA-cloning rather than i-MAXII™.
QRNA was extracted with easy-spin, and then cDNA was synthesized with Maxime RT. PCR was performed using i-MAXTM II, but PCR efficiency is poor.
AThank you for using our products. After synthesizing the cDNA, if the amount of template is large during PCR, it may cause PCR inhibition. Therefore, you can get better results if you run the experiment by reducing the amount of template. 
QThe fidelity of i-MAXII™ is high on the homepage. What is the error rate compared to i-pfu™?
AThe error rate of i-MAXTMII is 1.8X10-8. For i-pfu™, it is 1X10-6.
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