ABI Sanger Sequencing of Avian Clock genes to elucidate markers for Migration Phenology

Louis-Stéphane Le Clercq, Desire Lee Dalton, Antoinette Kotze, J. Paul Grobler

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Abstract

This protocol follows up on "PCR Amplification of Clock genes with EmeraldAmp® GT PCR Master Mix in Avian species" and is intended to provide the next steps used in the sanger sequencing of the produced amplicons. This protocol uses ABI BigDye reagents (but suitable alternatives exist). The same primers used to produce the PCR products, of the respective clock genes, are used individually in a forward and reverse sequencing reaction. Cycling conditions mimic those used for PCR. Sequencing reactions are purified and subsequently resolved on an ABI Genetic Analyzer. The sequence read data was used in a BLAST search and confirmed to be the genes and regions of interest for all tested species.
Original languageEnglish
JournalProtocols.io
DOIs
Publication statusPublished - 1 Nov 2023

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