This kit is a ready-to-use 2× premixed solution containing High-Fidelity DNA Polymerase, dNTPs, and the buffer system carefully optimized for amplification of fragments within 20 Kb, with an extension rate of 10 sec/kb. Its fidelity is 83 times that of Taq DNA polymerase and 9 times that of Pfu DNA polymerase. It is suitable for complex template amplification, and the amplified product has a blunt end.

This product is fast and simple, highly sensitive, specific and stable. It can amplify fragments with a GC content of 30% to 70%. It can amplify whole blood samples that have not been extracted with nucleic acids. It is suitable for blood preserved with EDTA, citrate or heparin, as well as dried blood spot cards, which is faster and more flexible. In addition, this product also contains a specific protective agent, so the premix can maintain stable activity after repeated freezing and thawing. All reagent components provided in this product have undergone strict quality inspection to ensure the excellent performance and batch-to-batch stability of the product to the highest extent. This reagent is suitable for different multiplex PCR needs. It can be used for gel electrophoresis to identify multiple products of fragments of different sizes within 20 pairs, and can also be used for enrichment of hundreds of or more amplicons.

Features

Easy to operate: ready-to-use 2× premixed solution

High fidelity: 83 times the fidelity of Taq enzyme

Long fragment amplification ability: high efficiency amplification within 40 kb

Fast amplification speed: ＜10 kb, 5 sec/kb；≥10k, 10 sec/kb

High sensitivity: amplify as low as 100 pg template

Broad Compatibility: Suitable for complex samples, GC-rich/poor systems(20-80%), and multiplex amplification.

Components

Application

Routine polymerase chain reaction (PCR); Multiplex PCR; Fast PCR; High-fidelity PCR

Figures

A. Long fragment amplification capability

Figure 1. Long fragment amplification test

Using human genomic gDNA as template, 17228ES and supplier K* and supplier V* were used to synchronously amplify 1-40 kb fragments.

Input DNA 50 ng, extension time: 10 sec/kb for 10 K and above, 5 sec/kb for below 10 Kb.

Marker: Yeasen 10510ES, GendON 50 kb large-range DNA Ladder.

Loading volume: 4 μL each after dilution 10 times

Conclusion: 17228ES can amplify genomic fragments up to 40 kb, and the extension speed is fast.

B. Broad Compatibility:

Figure 2. Compatibility Testing

GC compability: Complex GC content of 20%~80% can be amplified;

Using the reverse transcription product of 293 RNA as a template, 17228ES can effectively amplify cDNA of 1 Kb~14 Kb;

Direct PCR: Using 17228ES to amplify E. coli liquid, the input volume can reach 12.5%, and the amplification specificity is good, no tailing, and no non-specific bands.

C. High stability

Figure 3. Stability test

Placed at 37℃ for 2 weeks, 4℃ for 4 weeks, using 0.1 ng and 1 ng human gDNA to amplify 500 bp fragments, using 50ng genome to amplify 2 Kb , 10 kb and 40 kb DNA fragments, and setting the extension time to 5-10 sec/kb.

Result: After placement, the amplification performance did not change

Figure 4. Multiplex amplification effect (human 7-plex amplification, 3 kb-10 kb)

Sample: human gDNA, Input DNA 30 ng. Reaction conditions: 25 μL reaction system, the panel amount is 0.5μl-2μl, PCR amplification 35 cycle, Extension speed: 68℃ 120 sec.

The marker sizes are 3 kb, 4 kb, 6 kb, 7 kb, 9 kb and 10 kb.