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

Abstract

ABSTRACT. 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.

An Easy, Quick and Cost Effective Method of High Quality DNA Extraction from Mungbean [Vigna radiata (L.) Wilczek] Without Liquid Nitrogen

“…The need of liquid nitrogen during grinding step has been eliminated by many workers using soft tissues such as flower petals or young leaves (Khan et al, 2004), cold and heat shock treatments, (Biswas and Biswas, 2011) alcohol fixation (Sharma et al, 2003;Sharma et al, 2010). (Duan et al, 2015;Ibemhal et al, 2015). Here, we developed an easy, fast, inexpensive method to isolate high quality genomic DNA from mungbean eliminating the use of hazardous chemical phenol and expensive reagents like proteinase K and liquid nitrogen.…”

Comparison of Methods for Genomic Deoxyribonucleic Acid (DNA) Extraction Suitable for Whole-Genome Genotyping in Traditional Varieties of Rice

“…The rice DNA extraction methods typically use seedlings or leaves and require fine grinding of the tissue materials using liquid nitrogen. Different methods of DNA isolation have been reported in many studies (Ahmadikhah, 2009, Aliyu et al, 2013Fouladvand et al, 2013 andDuan et al, 2015). references Ahmadikhah, A.…”

Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks

“…Although efficient DNA extraction procedures and commercial kits are readily available for rice leaves as the starting material, very few studies have been conducted to establish DNA fingerprinting protocols for rice caryopses and flour (Mutou et al, 2014). Two protocols have been established for DNA extraction from rice seeds by Duan et al, (2015) and from half-cut rice seeds of the varieties Nipponbare (O. sativa Japonica) and Junyou522 (O. sativa Indica) by Liang et al, (2016). However, none of these studies used common varieties found in Sri Lanka.…”