Shotgun sequencing of Drosera capillaris: whole plant after chloroplast enrichment and multiple displacement amplification.
The plant sample was enriched for chloroplasts using the Minute Chloroplast Isolation kit (Invent biotechnologies, Plymouth, MN). DNA was then extracted using the DNeasy Plant Pro kit (Qiagen, Redwood City, CA). Multiple displacement amplification was performed using the REPLI-G kit (Qiagen). The amplified DNA was digested with Endonuclease T7 (New England Biolabs, Ipswich, MA) to reduce DNA structures and purified using Mag-Bind TotalPure NGS beads (Omega Bio-tek, Norcross, GA). The purified DNA was randomly sheared into shorter fragments, end-repaired, A-tailed, and further ligated with Illumina adapters. The resulting fragments with adapters were size selected, PCR-amplified, and purified with magnetic beads. The library was quantified through Qubit and qPCR, and size distribution detected with fragment analyzer. The quantified library was sequenced on an Illumina instrument.