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Here is a zipped file containing amplification and annealing protocols for the MJ Opticon real time PCR machine. If you are using this machine, the files may be used directly. The annealing files are named for the desired annealing temperature (usually set at Tm - 3°C). If you are using another machine, please open the files with a spreadsheet or text program to see a model for creating your own programs. To avoid melting the reference wells, it is important to choose the gradient temperatures carefully so that all the reference wells remain below the annealing temperature, and it is important to specify a "block" rather than "sample" thermostat setting.

Machine protocols (zip)

Last updated September 5, 2006