Courtesy of Redasoft

Redasoft's Visual Cloning 2000 includes primer design tools.

Courtesy of Sigma-Aldrich

Direct loading of PCR products onto an agarose gel using Sigma-Aldrich's REDTaq.

The art of PCR isn't hard to master. An abundance of products, ranging from relatively low-cost reagents to sophisticated optimization software, exists to meet most, if not all, PCR challenges. This profile looks at commonly used additives, PCR optimization kits and protocols, software tools for PCR enhancement, and hot-start polymerases. Other thermostable polymerases will be reviewed in the Sept. 3, 2001, issue of The Scientist.

The key to an optimal PCR experiment is primer design. In general, primers should be between 18 and 24 nucleotides long and have a GC content between 40 and 60 percent. Ideally primers should contain a relatively GC-rich 5'-end and an AT-rich 3'-end (i.e., containing three A or T residues in the last five bases)....

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