When using traditional high-fidelity enzymes for PCR, it's inevitable to encounter some thorny issues. For instance, the fidelity may not be sufficient, leading to mutations or deletions in the target bases even after sequencing the amplified fragments. The amplification time is also relatively long; sometimes, amplifying a 5kb fragment can take nearly 2 hours. Now, XiaoYi introduces a rapid high-fidelity PCR to solve your troubles! -- Hieff Canace™ AdvanceFast PCR Master Mix (Ultra-fast High-fidelity PCR Premix).
The 2× Hieff Canace™ AdvanceFast PCR Master Mix (With Dye) contains a pre-added electrophoresis indicator, allowing the PCR products to be directly used for electrophoresis. The amplified products have blunt ends. This high-fidelity enzyme mix offers the advantages of being fast, simple, highly sensitive, strongly specific, and stable. In the reaction system, only primers and templates need to be added. Additionally, the product also contains a specific protectant, which ensures that the high-fidelity PCR premix maintains stable activity even after repeated freezing and thawing.
Product Name
2× Hieff Canace™ AdvanceFast PCR Master Mix (With Dye)
Product Features
- High Fidelity: Low mismatch rate, with a base mismatch ratio of 231/300,000 in high GC mutation-prone sequences;
- Fast Amplification Speed: Extension rate as fast as 5 seconds per kb;
- Wide Applicability: Capable of covering genes with 20-80% GC content, tolerates mouse lysates and 20% blood;
- Good Stability: Can be stored at 37°C for 7 days without affecting product performance;
- Fast and Convenient: Simply add primers and template for amplification, comes with loading dye for direct electrophoresis.
Product Performance Demonstration
High Fidelity, Low Mismatch Rate
Using different species' genomes as templates, six GC-rich fragments that are highly prone to base mutations/losses were selected for amplification. The respective PCR products were cloned into vectors, and 100 single clones for each sequence were picked for sequencing to confirm base mismatch/mutation occurrences. The results showed that the rapid high-fidelity PCR master mix (10164ES) had a superior rate of base mismatch/mutation compared to domestic and international brand reagents.
The extension time can reach up to 5 seconds/kb
Using the rapid high-fidelity enzyme premix (10164ES) and Product T, different length fragments (0.4-10 kb) were amplified from 100 ng of human genomic DNA template at an extension rate of 5 sec/kb, with an extension time required to be 5 sec/kb. The results showed that the rapid high-fidelity enzyme premix (10164ES) can amplify fragments within 10kb at an extension rate of 5 sec/kb.
Can amplify gene fragments with different GC content
Using the rapid high-fidelity enzyme premix (10164ES) to amplify gene fragments with 23-76% GC content from human genomic DNA templates, with the extension time set at 5 sec/kb. The results showed that the product has good amplification specificity and is capable of amplifying fragments with different GC contents.
Capable of amplifying gene fragments with varying GC content
Using the rapid high-fidelity enzyme premix (10164ES) to amplify fragments of varying lengths from different samples, with an extension time set at 5-10 seconds per kilobase. The results indicate that this product exhibits excellent amplification specificity and is compatible with the amplification of different fragments from various templates.
Note: 1. Mouse genomic DNA 1 kb; 2. Mouse genomic DNA 2 kb; 3. Mouse genomic DNA 6 kb; 4. Corn genomic DNA 1 kb; 5. Rice genomic DNA 2 kb-A; 6. Rice genomic DNA 2 kb-B; 7. Human 293 cell cDNA 1 kb; 8. Human 293 cell cDNA 2 kb; 9. Human 293 cell cDNA 3 kb; 10. Human 293 cell cDNA 12 kb; 11. E. coli broth 8 kb; 12. E. coli broth 16 kb; 13. Rice genomic DNA 10 kb-A; 14. Rice genomic DNA 10 kb-B; 15. Rice genomic DNA 10 kb-C; 16. Rice genomic DNA 20 kb-A; 17. Rice genomic DNA 20 kb-B; 18. Human blood direct amplification 3 kb.
High detection rate of template amplification across different species
Using the rapid high-fidelity enzyme premix (10164ES), different GC content fragments ranging from 0.5-10 kb in Arabidopsis, rapeseed, and potato were amplified, with an extension time of 5 sec/kb. The results showed that the rapid high-fidelity enzyme premix (10164ES) can ensure successful amplification.
Using the rapid high-fidelity enzyme premix (10164ES), random sequences (randomly selected) from Arabidopsis, rapeseed, fruit flies, Caenorhabditis elegans, and zebrafish, with fragment sizes ranging from 0.5-10 kb and covering 20-80% GC content, were amplified. The extension time was set at 5 sec/kb. The results showed that the rapid high-fidelity enzyme premix (10164ES) has a high detection rate.
Good stability
Using the rapid high-fidelity enzyme premix (10164ES) that has been heat-accelerated stored at 37℃ for 3 days and 7 days, respectively, to amplify 3 kb and 5 kb DNA fragments from 100 ng of human genomic DNA, with an extension time set at 5 sec/kb. The results show that the product has good stability.
Customer Trial Feedback
Animal gDNA-120bp-50%GC
Customer evaluation: The PCR process is fast, the bands are appropriate, and can be directly loaded into agarose gel electrophoresis. The bands of N brand are biased upwards, which may be due to the influence of its own buffer, and since no loading buffer was added, this product can replace the high-fidelity enzyme in my current experimental protocol.
Plant cDNA-729bp-45.13%GC
Customer evaluation: Compared to the T brand, the cloning bands are clearer and more uniform, and the entire PCR process is faster. There is no need to add extra dye when running the gel, which is very convenient and time-saving.
Microbial gDNA-1158、1953bp-58.8%、52.9%GC
Customer evaluation: Compared to the V brand, the effect is comparable, but personally, I find it even better.
Plant cDNA-2000bp-40%GC
Customer evaluation: The effect is very good, and the strip is clear.
High-Fidelity PCR Selection Guide
|
Product Positioning |
Product Name |
Cat# |
|
5s/kb, ensures fidelity and speed (With Dye) |
2× Hieff Canace™ AdvanceFast PCR Master Mix (With Dye) |
10164ES |
|
5s/kb, ensures fidelity and speed (Without Dye) |
2× Hieff Canace™ AdvanceFast PCR Master Mix |
10163ES |
|
83×Taq, high detection rate (With Dye) |
2×Hieff Canace™ Plus PCR Master Mix(With Dye) |
10154ES |
|
83×Taq single enzyme + Buffer separate |
Hieff Canace™ Plus High-Fidelity DNA Polymerase |
10153ES |