Generating Pyrosequencing templates from PCR amplicons is fast and straightforward with the PyroMark vacuum workstation. These buffers and solutions are specifically designed for this purpose. PyroMark Binding Buffer facilitates binding of biotinylated PCR amplicons to streptavidin-coated Sepharose beads. These beads are then immobilized with the vacuum tool of the workstation. PyroMark Denaturation Solution gently separates the complementary strand from the biotin-tagged strand destined to become the Pyrosequencing template, and PyroMark Wash Buffer ensures that only the biotin-tagged, single-stranded DNA remains immobilized to the vacuum tool. Finally, when the single-stranded DNA is released into reaction vessels containing the Pyrosequencing primer, hybridization of primer and template is enhanced by PyroMark Annealing Buffer.