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

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2023-06-14 DOI:10.1002/aps3.11529

Jonathan Selz, Nicolas R. Adam, Céline E. M. Magrini, Fulvia Malvido Montandon, Sven Buerki, Sebastian J. Maerkl

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引用次数: 3

Abstract

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.

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一种利用微针片进行植物调查和监测的现场快速植物DNA提取方案

提出了一种利用微针快速提取植物DNA的新方案，该方案可支持植物调查、分类学和系统学。该方案可以在实验室技能和设备有限的情况下在现场进行。通过测序和BLAST分析与QIAGEN自旋柱DNA提取结果进行比较，验证了该方案。方法与结果对13种植物进行了两组DNA提取:(i)用定制的聚合物微针贴片在新鲜叶片上打孔以恢复基因组DNA，或(ii) QIAGEN DNA提取。三个质体(matK, rbcL和trnH-psbA)和一个核糖体(ITS) DNA区域扩增并使用Sanger或纳米孔技术测序。该方法将提取时间缩短至1 min，并获得与QIAGEN提取相同的DNA序列。结论该方法快速简便，适用于纳米孔测序，适用于基于dna的高通量物种鉴定和监测等多种应用。

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