Director™ Cloning System

Director™ Technology for Rapid Streamlined Cloning of Any Gene Sigma's Director Universal PCR system is an ExoClone™ technology-based solution that provides a rapid, universal and directional method to facilitate the streamlined cloning of large numbers of PCR products. This technology does not rely on base-recognition enzymes and thus overcomes problems of internal cutting sites and rearrangements associated with standard cloning methods. In addition, Director minimizes extra amino acids carried along in the cloning process compared to other methods. Overview of Method Products

Features and Benefits:

• Universal - Streamlined cloning of numerous genes without concerns of internal cutting sites or rearrangements
• Directional - Use of Exoclone technology provides new method of generating cohesive ends
• Rapid - Simple procedure increases throughput while decreasing hands-on time
• Versatile - Prepare your own vector or use our Director-Ready™ vectors
• High efficiencies - Typical cloning efficiency using the Director-Ready vectors is greater than 90%

Overview of the Director™ Cloning Method

The Director system uses a specially formulated dNTP mix containing thionucleotides dATPαS and dGTPαS. After PCR using gene-specific primers (with appropriate 5 base overhangs), cohesive 5'-termini of the amplicon are produced by Exonuclease III digestion instead of being generated by traditional restriction enzymes. Exonuclease III degrades linear duplex DNA sequentially from 3' termini, and incorporation of dA/GTPαS into the amplicon provides a convenient method for protecting the amplicon from overdigestion. The PCR products are then ready for directional ligation into any expression vector previously digested with 5' overhang producing enzymes (i.e., Director-Ready vectors). The complete system includes the universal PCR kit and either of two Director-Ready vector kits.

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