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Oil-Free PCR Using the Hot Top Assembly

Hot Top Assembly now available for Robocycler 96 temperature cyclers

Michelle Cayouette Jane Moores Michael Steege Jason Capodanno Michael Rahilly
Stratagene Cloning Systems, Inc.

With the introduction of the Hot Top Assembly for RoboCycler 96 temperature cyclers, researchers can begin running oil-free PCR amplifications. Each Hot Top Assembly for RoboCycler 96 cyclers is supplied with a software upgrade card, which provides more accurate temperature control. Hot Top Assemblies are available as options on newly purchased RoboCycler temperature cyclers.# Individual units that can be added to existing RoboCycler 40 or 96 units are also available. Using both gradient and single-temperature RoboCycler 96 models, we show the yield and specificity of PCR products to be equivalent before and after the addition of the Hot Top Assembly and software upgrade. Furthermore, PCR results at a given annealing temperature were reproducible whether the annealing block was run in gradient or single-temperature mode, an important feature for optimizing annealing temperatures. When used for small-volume cycle sequencing experiments, the Hot Top Assembly did not affect results adversely, and gel loading was quicker and easier without oil or wax overlays on reactions.

Stratagenes line of RoboCycler temperature cycler currently includes four models. The RoboCycler 40 temperature cyclers accommodate up to forty 600-l individual PCR tubes; the RoboCycler 96 units are high-throughput models that accommodate either a 96-well plate or ninety-six 200-l individual or strip tubes. Both the RoboCycler 40 and 96 temperature cyclers are available in either gradient or nongradient models. Gradient models facilitate PCR optimization by allowing researchers to simultaneously test multiple annealing temperatures. All members of the Rob
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