Optimizing efficient PCR?amplifiable DNA extraction from herbarium specimens

Premise The objective of this study was to optimize an existing DNA extraction protocol for recalcitrant plant taxa obtain high-quality from preserved herbarium tissue suitable downstream PCR applications. Methods and Results Leaf 30 diverse species obtained the U.S. National Arboretum Herbarium. Our previous (Gouker et al., 2020, Applications in Plant Sciences 8: e11403) improved by use 10X Tris-EDTA buffer, addition polyvinylpolypyrrolidone, omission sample heating during homogenization, resulted total yields ranging 60–2460 ng. Optimized amplification using universal primers ITS-p3/u4 region P6 loop trnL (UAA) chloroplast intron successful most specimens. Conclusions This protocol, which is simple, fast, uses standard laboratory-grade chemicals, specimens that comparable quality commercially available kits, sufficient quantity other