Establishment of simple genomic DNA extraction method for marker assisted selection in lettuce
2019
Seki, K. (Nagano Prefecture Vegetable and Ornamental Crops Experiment Station (Japan))
To use DNA markers for plant breeding, it is necessary to quickly extract genomes for PCR from multiple samples. If the number of samples is small, it is possible to reliably extract the genome to be used for PCR by using a proven commercial genomic extraction kit, but when the number of samples is large, experiment labor and cost will be a problem. By applying the glass fiber filter paper method which can extract the genome by utilizing the property that DNA is easy to bind to silicon dioxide, a simpler method of genome extraction was examined. Lettuce, from which it is difficult to extract the genome, was used for the test crop. As a result, it was confirmed that genome can be quickly extracted from multiple samples by placing the glass fiber filter paper in a pipette tip and pipetting the crude extract of genome and the two washing solutions several times respectively. PCR using genomes extracted by this method revealed that amplified products can be stably obtained using KOD FX. In this study, I established a method to rapidly extract the genome of lettuce used for PCR and stably obtain PCR products.
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