2X PCR Master mix Solution (i-StarMAX™ GH)

2x PCR Master mix type with high sensitivity, specificity and amplification, including Antibody based-Hot Start function and Proof-reading function

• Application of i-StarMAX™ GH DNA Polymerase
– Antibody-based hot-start implementation enhances sensitivity / specificity
– Proof-reading function
– Possible to amplify fragments over 20 Kb
• Ready-to-use
– All reagents required for PCR reaction are contained in solution in one tube.
– If template, primer and D.W are added, PCR will be performed immediately.
• PCR reaction volume control
• Optimal product for long PCR that conducts PCR of complex structure using human genomic DNA as a template, or studies the treatment of diseases and diseases.

Theoretically, the length of a fragment that can be amplified using conventional Taq is less than about 5 Kb and the ideal length is less than 3 Kb. Longer fragments longer than 10 Kb can be amplified by standard PCR techniques, but longer fragments have lower amplification efficiency and, because of the difficulty of obtaining stable results, to achieve long PCR amplification efficiency and stable results, There are many known PCR techniques that can amplify from 20 Kb to 40 Kb. The feature of Taq used for the amplification of longer fragments is known to have the greatest influence on the detection of pfu Taq isolated from P. furiosis with proof-reading function.

Proof-reading is defined as the template template is synthesized when a base that does not form a base pair correctly is inserted into the DNA strand of the clone, so that the base is removed to ensure accurate DNA replication. As mentioned earlier, unlike Taq DNA Polymerase, pfu DNA Polymerase has a function to stop synthesis automatically when a wrong base sequence is inserted into a template in PCR and to find an appropriate base sequence. Accuracy of PCR is higher than that of normal Taq But it is possible only in Long PCR and it takes long time to perform PCR. Therefore, due to advantages and disadvantages of Taq DNA Polymerase and pfu DNA Polymerase, Taq DNA Polymerase and pfu DNA Polymerase are blended at a certain ratio to complement each other to improve PCR accuracy and amplification rate.

2X PCR Master Mix Solution (i-StarMAX™ GH) uses i-StarMAX ™ DNA Polymerase, which is blended with Taq DNA Polymerase and pfu DNA Polymerase. Therefore, it has high amplification efficiency from short to long fragments. It is developed as master mix type to provide convenience. 2X PCR Master Mix Solution (i-StarMAX^TM GH) is designed to be used with template, primer and DW only. In addition, gel loading dye is included. The PCR products amplified using this product are highly accurate and stable, and are highly reliable in subsequent experiments such as cell ligation and cell transformation through TA cloning or restriction enzyme treatment, and cDNA library construction through these methods can be used.

  Applications

  Kit Contents

  Technical Data

Amplification efficiency under various conditions

Based on various PCR amplification conditions, we found that the sensitivity and amplification rate of 2X PCR Master Mix Solution (i-StarMAX ™ II) were superior to those of existing products.

Fig 1. PCR amplification of 1.3/2.7/4.5/20 Kb with 2X PCR Master mix Solution (i-StarMAX™ II) and 2X PCR Master mix Solution (i-StarMAX™ GH)

[ 1.3 Kb ] Lane M, SiZer™-100 DNA Marker Solution(Cat. No. 24073); lane N, Negative Control
Lane 1, 100 ng K562 gDNA; lane 2, 10 ng K562 gDNA; lane 3, 1 ng K562 gDNA; lane 4, 100 pg K562 gDNA; lane 5, 10 pg K562 gDNA; lane 6, 1 pg K562 gDNA; lane 7, 100 fg K562 gDNA 

[ 2.7 Kb ] Lane M, SiZer™-1000 DNA Marker Solution(Cat. No. 24074); lane N, Negative Control
Lane 1, 100 ng K562 gDNA; lane 2, 10 ng K562 gDNA; lane 3, 1 ng K562 gDNA; lane 4, 100 pg K562 gDNA; lane 5, 10 pg K562 gDNA; lane 6, 1 pg K562 gDNA; lane 7, 100 fg K562 gDNA 

[ 4.5 Kb ] Lane M, SiZer™-1000 DNA Marker Solution(Cat. No. 24074); lane N, Negative Control
Lane 1, 100 ng 5F plasmid DNA; lane 2, 10 ng 5F plasmid DNA; lane 3, 1 ng 5F plasmid DNA; lane 4, 100 pg 5F plasmid DNA; lane 5, 10 pg 5F plasmid DNA; lane 6, 1 pg 5F plasmid DNA; lane 7, 100 fg 5F plasmid DNA 

[ 20 Kb ] Lane M, SiZer™-1000 DNA Marker Solution(Cat. No. 24074); lane N, Negative Control
Lane 1, 10 ng λDNA; lane 2, 1 ng λDNA; lane 3, 100 pg λDNA; lane 4, 10 pg λDNA; lane 5, 1 pg λDNA ; lane 6, 100 fg λDNA; lane 7, 10 fg λDNA