Abstract

A simplified genomic DNA extraction protocol for pre-germination genotyping in rice

Author(s): Y.B. Duan, F.L. Zhao, H.D. Chen, H. Li, D.H. Ni, P.C. Wei, W. Sheng, J.T. Teng, A.M. Zhang and J.P. Xue

Genotyping is a critical step for molecular markerassisted selection in rice. Rice genomic DNA samples for genotyping are typically isolated from living tissues such as seedlings. This requires the germination of all candidate seeds and extraction of DNA from the seedlings. Currently, an ideal individual is selected from a very large number of plants, which is time- and labor-consuming, requiring several transplantations of materials and sampling processes. In this study, we developed a simplified genomic DNA extraction protocol in rice by using amylase to treat half-seeds. The yields of genomic DNA from a half-seed of Indica and Japonica rice were greater than 203.8 ± 32.5 and 143.2 ± 25.5 ng, respectively, and the 260/280 nm absorbance ratio was 1.75-2.10. The DNA was confirmed to be sufficient for polymerase chain reaction amplification and can be used in a markerassisted selection program.

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