Microsatellite Multiplexing in Fish

Abstract
Microsatellite multiplexing is a powerful technique that can increase the productivity of genetic studies in fisheries biology. We review multiplexing methods and present an optimized and detailed protocol for microsatellite multiplexing that is specifically tailored for use with radioisotopes. The protocol can significantly reduce the cost associated with microsatellites and provides high polymerase chain reaction (PCR) fidelity and band resolution. Comparing three radioisotopes, we find that end labeling with 33P provides the highest resolution. We also present a quick and inexpensive DNA isolation protocol that is successful with fish larvae. Finally, we find that PCR fidelity depends on the quality of the DNA template, and we therefore review preservation and isolation methods specific to various fish tissue types. Together, these microsatellite multiplexing and DNA isolation protocols can significantly reduce the time and expense associated with genetic analyses in fish.

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