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Pringle Lab, Grossman Lab 2018. Symbiodinium cp23S RFLP and sequence analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.qyjdxunCopy Citation Copied
URL: https://dx.doi.org/DOI:10.17504/protocols.io.qyjdxun
Authors: Pringle Lab, Grossman Lab
Group: Aiptasia-Symbiodiniaceae Model System
Summary: This protocol describes how to perform PCR on extracted Symbiodinium DNA to amplify a fragment of the chloroplast 23S rRNA gene. It is based on the method describes in Pochon et al., 2006. The protocol then explains how to perform an RFLP (restriction fragment length polymorphism) analysis on the PCR products as well as eitherusing them directly for sequence analysis by Sanger sequencing or cloning them into a plasmid back bone to separate genes from mixed Symbiodinium populations and perform the sequence analysis on several clones per original sample.The attached file contains reference sequences of the following Symbiodinium strains: Clade A strains SSA01, SSA02, SSA03; SSB01 (clade B), SSB01del6, SSB01del136, SSE01 (clade E) and A001 (Clade D).
Affiliations: Stanford University, Carnegie Institution for Science - Department of Plant Biology
Version: 1
Publication Date: 2018
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