Development of Simple Sequence Repeat (SSR) and Morphological Markers to Identify Jaboticaba Cultivars

Jaboticaba is an evergreen fruit tree in Myrtaceae family, native to the southern region of Brazil, Paraguay and the northern region of Argentina. We carried out the microsattelite capture sequencing technique that enable to detect simple sequence repeat (SSR) region of specific motif using high-throughput sequencing teghnology to create an extensive SSR catalog for this plant. Briefly, 1 microgram of the DNA was fragmented by shearing to an average fragment size of 500 bp using an adaptive focused acoustics sonicator (Covaris). A pre-enriched library was constructed from the fragmented DNA using the NEBNext Ultra DNA Library Prep kit for Illumina (New England Biolabs). A customized biotinylated SSR probe (GA)10 was applied to enrich the library with SSR region of (GA)n-motif. After washing by SSC buffer, the SSR-enriched library was finally amplified by 15 cycles of high-fidelity PCR. The library was sequenced using 2 x 300 bp paired-end sequencing using a MiSeq (Illumina).