Robust PCR amplification of GC-rich human genomic DNA
A serial dilution in duplicate of human genomic DNA (1 μg, 200 ng, 100 ng, 50 ng, 25 ng and 12.5 ng, 1-6 respectively) was used with primers and Taq DNA Polymerase to amplify a 450 bp fragment of the human myc gene. The section of gene used has 61% GC content and was selected to demonstrate the high yield and sensitivity of the Taq DNA Polymerase, making it the perfect choice for the amplification of complex PCR templates.
Taq PCR Buffer, 5x, MDX002
An optimized, novel buffer system, containing dNTPs, MgCl2 and enhancers, to give superior amplification, making it the perfect choice for complex templates.
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Description
Taq PCR Buffer is a combination of the latest advances in buffer chemistry together with enhancers, stabilizers, dNTPs, and MgCl2 at optimal concentrations. Taq PCR Buffer has been designed for superior fast amplification across a range of templates.
Specifications
| Description | An optimized, novel buffer system, containing dNTPs, MgCl2 and enhancers, to give superior amplification, making it the perfect choice for complex templates. |
| Concentration | 5x |
| Appearance | Clear, colorless solution |
| Application | PCR, qPCR |
| Sample type | cDNA, DNA |
| Presentation | 1 vial |
| Storage | -20 °C |
| Mix stability | See outer label |
| Functionality | PCR with a dilution series of human genomic DNA, single, distinct band was observed |
| DNA Contamination | None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. |
| DNase Contamination | No detectable degradation |
产品资料
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FAQs: Taq DNA Buffer
Taq DNA Buffer has been validated for use with a broad range of PCR templates including DNA extracted from human, animal and plant samples, making it the ideal choice for the following applications:
• End-point PCR testing
• High-yield PCR testing
• Fast PCR testing
• Multiplex PCR testing
For qPCR we recommend the use of Fast qPCR Buffer, 4x (MDX033) as it has been optimized for this application.
This depends on the polymerase used as well as the length of the amplicon and the complexity of the template. For fast PCR, with Taq HS DNA polymerase (MDX008), the initial denaturation is 1 minute, but with low complexity template such as plasmid DNA, an annealing of 5 seconds and extension time of 10 seconds is sufficient for amplicons up to 1 kb. In order to find the fastest optimal condition, for more complex or longer amplicons, we suggest incrementally increasing the extension time successively up to 30 s/kb.
Decreasing the template concentration will help to minimize the concentration of inhibitors, the amount of DNA polymerase and primer can also be increased. In some cases, you may also have to add a pre-treatment step if the DNA is not accessible.
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