TY - JOUR
T1 - A Field‐Capable Rapid Plant DNA Extraction Protocol Using Microneedle Patches for Botanical Surveying and Monitoring
AU - Selz, Jonathan
AU - Adam, Nicolas R.
AU - Magrini, Céline E. M.
AU - Montandon, Fulvia Malvido
AU - Buerki, Sven
AU - Maerkl, Sebastian J.
N1 - Publisher Copyright:
© 2023 The Authors. Applications in Plant Sciences published by Wiley Periodicals LLC on behalf of Botanical Society of America.
PY - 2023/5/1
Y1 - 2023/5/1
N2 - Premise: A novel protocol for rapid plant DNA extraction 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 sequencing and comparing the results with QIAGEN spin-column DNA extractions using BLAST analyses. Methods and Results: Two 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, or (ii) QIAGEN DNA extractions. Three plastid ( matK , rbcL , and trnH-psbA ) and one nuclear ribosomal (ITS) DNA regions were amplified and sequenced using Sanger or nanopore technology. The proposed method reduced the extraction time to 1 min and yielded the same DNA sequences as the QIAGEN extractions. Conclusions: Our drastically faster and simpler method is compatible with nanopore sequencing and is suitable for multiple applications, including high-throughput DNA-based species identifications and monitoring.
AB - Premise: A novel protocol for rapid plant DNA extraction 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 sequencing and comparing the results with QIAGEN spin-column DNA extractions using BLAST analyses. Methods and Results: Two 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, or (ii) QIAGEN DNA extractions. Three plastid ( matK , rbcL , and trnH-psbA ) and one nuclear ribosomal (ITS) DNA regions were amplified and sequenced using Sanger or nanopore technology. The proposed method reduced the extraction time to 1 min and yielded the same DNA sequences as the QIAGEN extractions. Conclusions: Our drastically faster and simpler method is compatible with nanopore sequencing and is suitable for multiple applications, including high-throughput DNA-based species identifications and monitoring.
KW - DNA extraction
KW - field
KW - microneedles
KW - plant
KW - rapid
KW - sequencing
UR - https://scholarworks.boisestate.edu/bio_facpubs/772
UR - http://www.scopus.com/inward/record.url?scp=85161870599&partnerID=8YFLogxK
U2 - 10.1002/aps3.11529
DO - 10.1002/aps3.11529
M3 - Article
VL - 11
JO - Applications in Plant Sciences
JF - Applications in Plant Sciences
IS - 3
M1 - e11529
ER -