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ARS Home » Plains Area » Lincoln, Nebraska » Agroecosystem Management Research » Research » Publications at this Location » Publication #409033
Research Project: Management of Stable Flies to Improve Livestock Production

Location: Agroecosystem Management Research

Title: Crop DNA extraction with lab-made magnetic nanoparticles

Author
item Wang, Haichuan (john)
item ZHAO, XUEQI - University Of Nebraska
item TAN, LI - University Of Nebraska
item Zhu, Junwei - Jerry
item HYTEN, DAVID - University Of Nebraska

Submitted to: PLOS ONE
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/21/2023
Publication Date: 1/8/2024
Citation: Wang, H., Zhao, X., Tan, L., Zhu, J.J., Hyten, D. 2024. Crop DNA extraction with lab-made magnetic nanoparticles. PLOS ONE. 19(1):1-11. https://doi.org/10.1371/journal.pone.0296847.
DOI: https://doi.org/10.1371/journal.pone.0296847

Interpretive Summary: High quality DNA preparation from crops is challenge. The tradition DNA extraction method is time consuming, costly and not environment-friendly. In current study, by combination of magnetic nanoparticles and a commonly used extraction buffer, we successfully prepared quality DNA from three crops (soybean, wheat and maize). Downstream analysis confirmed its suitability in applied molecular breeding programs for desired traits selection at genome level.

Technical Abstract: A quick and inexpensive high-throughput method for isolating A quick and inexpensive high-throughput method for isolating genomic DNA from two different tissues of three crops; maize (Zea mays), soybean (Glycine max) and wheat (Triticum aestivum) was tested with the DNA isolation method. Combining CTAB extraction buffer and lab-made SA -coated magnetic nanoparticles, quality genomic DNA was isolated from leaf tissue (soybean, wheat and maize) and seed (soybean and wheat) in less than 2 hours with less steps than a standard CTAB extraction method. The single, clear band of DNA samples on 1.5% agarose gel demonstrated this method isolated high molecular weight DNA. The yield of the genomic DNA was 582-729 ng per 5 leaf discs or 216-1869 ng per seed in soybean, 2.92-62.6 ng per 5 leaf discs or 78.9-219 ng per seed in wheat, and 30.9-35.4 ng per 5 leaf discs in maize. The isolated DNA was tested with multiple molecular breeding methods and was found to be of sufficient quality and quantity for PCR and targeted genotyping by sequencing methods such as MIPs.