RDC1
 |  |  |
 |
Image
Useful Links & Tools
RDC1
Sigma
Director™ Universal PCR System
Description
| Application | The simple three step procedure (PCR, Exonuclease III digestion and rapid ligation/transformation) can be completed in one day. The typical cloning efficiency using this method is greater than 80%. |
| Features and Benefits | • Universal - PCR amplicon can be cloned into any expression vector • High Cloning Efficiency - Typically >80% • High Expression Efficiency - Typically >66% • High Fidelity - Long and accurate, hotstart enzyme generates amplicons up to 20 kb with fidelity up to 6.5× greater than standard Taq DNA polymerase • Fast - Simple three-step procedure allows completion in less than one day |
| Other Notes | The Director Universal Cloning System provides a simple, rapid and universal method to directionally clone PCR products into a vector cleaved with 5′ overhang-producing restriction endonucleases. |
| Principle | Directionality is achieved by pairing directionally designed PCR primers (e.g., containing restriction sites) with any appropriately digested plasmid. The kit contains an optimized nucleotide triphosphate mix, containing dATPαS and dGTPαS, that is used for the PCR step. After PCR, the cohesive 5′ termini of the amplicon are generated by Exonuclease III digestion instead of being generated by traditional restriction enzyme digestions. The dA/GTPαS that was incorporated into the amplicon during PCR protects it from over-digestion by Exonuclease III. The nucleotide mix in the kit is specially formulated so that the amplicon terminates at a statistically determined array of 3′ dA/GαS sites. PCR primers are designed such that the 5′ termini complement the 5′ overhangs of the predigested plasmid. |
| Legal Information | Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,079,352, 5,789,224, 5,618,711, 6,127,155 and claims outside the US corresponding to US Patent No. 4,889,818. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim (such as the patented 5′ Nuclease Process claims in US Patents Nos. 5,210,015 and 5,487,972), no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. |
| Director is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. |
Properties
| usage | sufficient for 25 PCR reactions |
| shipped in | wet ice |
| storage temp. | −20°C |
Safety
Components
| Kit component only | 10x AccuTaq™ LA DNA Polymerase Buffer 500 μL |
| Control PCR Template, 1ng/μl 10 μL | |
| Control RDC primer-R (with 5′ phosphorylation) 25 μL | |
| Control RDC primer-F (with 5′ phosphorylation) 25 μL | |
| ExoNuclease III, 100 units/μl 25 μL | |
| ExoClone™ 20x DNTP Mix 1 vl | |
| JumpStart™ REDAccuTaq™ LA DNA Polymerase, 1 unit/μl 62.5 μL | |
| Molecular biology grade water 500 μL |
