Abstract
The genus Lavandula comprises of several economically important lavender species that are mainly cultivated worldwide for essential oil production. Identification of lavender species and their cultivars has been a huge bottleneck in lavender industries due to lack of appropriate identification mechanisms. Recent advances in modern technologies would help to address these identification issues through development of potential molecular markers, including simple sequence repeats (SSRs). SSRs can be developed from specific species, and can be potentially used for related species, which lack the source sequences to develop species-specific SSRs. Here, we describe the guidelines and steps of identifying and analyzing SSRs from expressed sequence tag (EST) sequences of lavender species. We also detail the validation procedures of selected EST-SSRs in distinguishing source (donor) species as well as related species.
Keywords: EST-SSR, Lavandula, SSR, Lavender, Essential oil
Materials and Reagents
Equipment
Software
Procedure
Notes
Mature leaf tissue may contain large amounts of phenolic compounds that can co-purify with genomic DNA, and interfere with the subsequent PCR amplification reactions. Thus, extra caution must be taken on selection of tissue for DNA extraction. Young and fresh leaf tissues (Figure 2A) often give better DNA than mature and dry tissues.
Recipes
Acknowledgments
This protocol was adapted from the previously published study, Adal et al. (2015). This work was supported through grants and/or in-kind contributions to SSM by UBC, Genome British Columbia, Natural Sciences and Engineering Research Council of Canada, Agriculture and Agri-Food Canada and the BC Ministry of Agriculture (through programs delivered by the Investment Agriculture Foundation of BC).
References
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