ArticlePlant methods2025
A simplified low-cost and reliable plant genomic DNA extraction method for PCR-based genotyping and screening.
Article in Plant methods, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.
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Who cites it
2 citing papers in PubMed.
- Genotype-Specific Interactions Between Wheat and Rhizosphere Microbiome Under Drought Stress.Plants (Basel, Switzerland) · 2026Article
- Genome-wide data discovery of genetic variations in an advanced rice mutant line, NMR191, with blast resistance and early maturity traits.Data in brief · 2026Article
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Authors and funding
6 authors.
Funding
Abstract
backgroundExtraction of plant genomic DNA is a critical step for PCR-based genotyping, mapping, and breeding applications. Conventional CTAB protocols and commercial kits provide reliable DNA but are labour-intensive, costly, and generate substantial plastic waste. Simplified crude-extract methods are available, yet their performance is often compromised by PCR inhibition from salts and cellular debris. A rapid, low-cost, and high-throughput method is therefore needed for routine molecular applications.
resultsWe developed a single-tube DNA extraction protocol that eliminates supernatant transfers, thereby reducing handling errors, plastic consumption, and processing time. The method consistently produces DNA of sufficient yield and purity for PCR-based assays. Validation in wheat and wheat-wild relative introgression lines demonstrated robust amplification in KASP assays. Cross-species testing in maize, Arabidopsis, and tomato using two Tris-salt extraction buffers confirmed broad applicability, supported by NanoDrop and Qubit measurements. Freeze-dried and frozen tissue produced higher yields than fresh samples, confirming their suitability for high-throughput and large-scale studies.
conclusionsThis streamlined protocol provides a cost-effective, reliable, and scalable approach for extracting plant genomic DNA suitable for PCR-based genotyping, marker development, and diversity analysis. Its simplicity and throughput make it particularly valuable for breeding programmes, although it is not intended for applications requiring highly pure DNA, such as whole-genome resequencing.
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