Efficient and Robust Genomic DNA Isolation and Next-Generation Sequencing Library Preparation from Recalcitrant Wild Grape Species
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This protocol details the extraction of high-molecular-weight genomic DNA from grapevine tissues (wild and cultivated Vitis spp., including pathogen-infected samples) and the subsequent preparation of Illumina(R) whole-genome sequencing libraries using bead-bound Tn5 transposase. It is designed to overcome challenges from polyphenolic compounds and secondary metabolites in wild plants, providing a cost-effective workflow for large-scale population genomics. It includes recipes for buffers, incubation times, critical notes, and troubleshooting tips to maximize yield and library quality. Although designed for the grapevine DNA, this protocol is potentially applicable to other similar wild plant species
Highlights
Optimized CTAB-PTB DNA extraction protocol for field-collected wild plant tissues.
Effective removal of polyphenols and secondary metabolites associated with DNA using PTB.
Cost-effective Illumina Ⓡ DNA Prep library preparation using bead-bound Tn5 transposase (Tagmentation).
Scalable workflow suitable for large-scale population genomics in Vitis species.
Validated method for high-molecular-weight DNA and high-quality sequencing data.
