A field-capable rapid plant DNA extraction protocol using microneedle patches for botanical survey and monitoring

PremiseA novel protocol for rapid plant DNA extractions using microneedles is proposed, which supports botanic surveys, taxonomy and systematics. This protocol can be conducted in the field with limited laboratory skills and equipment. The protocol is validated by conducting sequencing and comparing results with Qiagen spin-column DNA extractions using BLAST analyses. Methods and ResultsTwo sets of DNA extractions were conducted on 13 species spanning various leaf anatomies and phylogenetic lineages: i) fresh leaves were punched with custom polymeric microneedle patches to recover genomic DNA ii) Qiagen DNA extractions. Three plastid (matK, rbcL, trnH-psbA) and one nuclear ribosomal (ITS) DNA regions were amplified, and Sanger or Nanopore sequenced. The proposed method reduced the extraction to 1 min and yielded the same DNA sequences as Qiagen. ConclusionsOur drastically faster and simpler method is compatible with Nanopore sequencing and suitable for multiple applications including high-throughput DNA-based species identification and monitoring.