High-throughput microsatellite detection using shotgun genomic circular consensus sequencing
Genomic DNA was isolated from the freeze-dried leaves using DNeasy Plant Mini Kit and eluted into 100 *L of different buffers. QIAGEN AE buffer was used in A. costaricensis and T. cumingiana, and QIAGEN EB buffer in C. insignis. DNA quality was checked using NanoDrop 2000, and dsDNA concentration was assessed using Qubit* 2.0 Fluorometer and Quant-iTTM dsDNA HS Assay Kits. Double-strained DNA of at least 30 ng/*L in a 50-*L volume from one tree of each species was used for PacBio 500 bp DNA library preparation following the manufacturer’s instructions. Four SMRT cells were running for each species on a PacBio RS sequencer using C2 chemistry