Rui Shi1, Dilip R Panthee2. 1. Department of Horticultural Science, North Carolina State University, Mountain Horticultural Crops Research and Extension Center, 455 Research Drive, Mills River, NC, 28759, USA. 2. Department of Horticultural Science, North Carolina State University, Mountain Horticultural Crops Research and Extension Center, 455 Research Drive, Mills River, NC, 28759, USA. dilip_panthee@ncsu.edu.
Abstract
MAIN CONCLUSION: A simple, low cost and safe method using homemade filter paper-based 96-well spin plates and homemade solutions was developed for high throughput plant DNA extraction to use in molecular marker analysis. A low cost and safe method was developed for high throughput extraction of plant DNA for molecular marker analysis. In this method, we describe a simple way to prepare 96-well spin plate using filter paper, a plant material product for DNA binding. Such filter paper-based spin plate can combine with homemade non-toxic buffers for high throughput extraction of plant DNA. We confirmed that filter paper is an efficient solid-phase DNA binding material and comparable to silicon-based glass fiber filters adopted in commercial DNA extraction kits, and that plant DNA extracted by this method can be readily used as template for PCR. The efficacy of this method was also fully demonstrated by molecular marker analysis in segregating populations of tomato. Due to greatly reduced expense compared to commercial kits, this method is of great value for small labs with limited resources.
MAIN CONCLUSION: A simple, low cost and safe method using homemade filter paper-based 96-well spin plates and homemade solutions was developed for high throughput plant DNA extraction to use in molecular marker analysis. A low cost and safe method was developed for high throughput extraction of plant DNA for molecular marker analysis. In this method, we describe a simple way to prepare 96-well spin plate using filter paper, a plant material product for DNA binding. Such filter paper-based spin plate can combine with homemade non-toxic buffers for high throughput extraction of plant DNA. We confirmed that filter paper is an efficient solid-phase DNA binding material and comparable to silicon-based glass fiber filters adopted in commercial DNA extraction kits, and that plant DNA extracted by this method can be readily used as template for PCR. The efficacy of this method was also fully demonstrated by molecular marker analysis in segregating populations of tomato. Due to greatly reduced expense compared to commercial kits, this method is of great value for small labs with limited resources.
Entities:
Keywords:
DNA extraction; Filter paper; High throughput; Inexpensive; Ph-3 gene; Safe; Solanum lycopersicum
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