Description
PCR optimisation kit for end-point applications. Includes everything needed to optimise PCR performance and achieve consistent, high-fidelity results for every DNA template.
The PCR PreMix Selection Kit includes the FailSafe PCR Enzyme Mix (24 uL), a blend of thermostable enzymes containing a 3′→5′ proofreading enzyme for high fidelity, and all twelve of the FailSafe PCR 2X PreMixes (100 uL each). 2X PreMixes contain dNTPs, buffer, different concentrations of MgCl2, and PCR Enhancer with betaine.
Simply add template, primers, and FailSafe PCR Enzyme Mix to each of the PreMixes and amplify to determine your optimal PCR conditions.
Key features
- PCR performance optimisation
- Amplification of high-GC templates and robust amplification of targets up to 20 kb in length
- Multiplex PCR compatible
- Fidelity at least 3x higher than Taq DNA polymerase.
- Extremely high sensitivity and specificity
Product information
The PCR PreMix Selection Kit is ideal for optimising PCR conditions and performance. The kit is multiplex PCR compatible, can amplify even the most difficult high-GC templates, and targets up to 20 kb in length.
Optimisation is done by adding a template, primers, and FailSafe PCR Enzyme Mix to each of the 12 FailSafe 2x PreMixes, followed by amplification. Results are analysed by agarose gel electrophoresis, which can indicate your best template/primer pair combination based on the 12 individual FailSafe 2X PreMixes.
Once optimal PCR conditions are determined; consistent and high-fidelity PCR results can be achieved through replication with the same template/primer and PreMix combination. The FailSafe PreMix Selection Kit is sufficient for 48 reactions (four optimization runs with each of the 12 total 2X PreMixes). Following optimization the FailSafe 2x PreMixes and Enzyme Mix can be purchased with the FailSafe PCR System as the bundled and affordable option, or, as individual components.
Applications
- End point PCR applications
- Cloning
- Sequencing
- Gene Expression
- Mutation Analysis
How It Works
Step 1
Start with the FailSafe PCR PreMix Selection Kit to optimise end-point PCR conditions.
Step 2
Analyse and determine which FailSafe PCR 2x PreMixes produce high-fidelity results, based on the template/primer pair combination.
Step 3
Purchase the post optimization FailSafe PCR System, which includes the Enzyme Mix plus your choice of 1, 2, or 8 out of the 12 PCR 2x PreMixes, OR, purchase the FailSafe PreMix and Enzyme components individually; and achieve reliable and consistent high-fidelity PCR results.
For a complete explanation of how FailSafe technology works, please read the manual.
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| Figure 1. Amplification of an 80%-85% GC-rich region of the human fragile X gene. PCR was performed using the FailSafe™ PCR System, schematically depicted above. Lanes A-L show the amplification products resulting from PCR using the 12 FailSafe PCR PreMixes. Lane M, molecular weight marker. In this experiment, optimal amplification was obtained with FailSafe PCR PreMix J. The size of the expected amplicon is indicated by an arrow. | Figure 2. Multiplex PCR of the human CFTR gene. The FailSafe™ PCR System amplified all five exons of the CFTR gene from as little as 1 ng of human genomic DNA. L |
