Abstract
The purity of DNA is one of the major factors affecting the success of Genomic studies. Nucleic acid isolation from polyphenol rich plants fail to produce good quality DNA or RNA as polyphenols adhere and interfere with DNA during isolation. An improvised, simple and inexpensive protocol has been developed for extracting genomic DNA from Mulberry (Morus spp.). The purity of the DNA as revealed by the ratios of absorbance at 260/280 nm (A 260/280) and 260/230 nm (A 260/230) was closer to 2.0. Genomic DNA analyzed for analytical applications like restriction digestion and PCR amplification with molecular markers viz., Inter Simple Sequence Repeats (ISSR), Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeat (SSR) primers further confirmed the purity of the DNA. A modified method of silver staining was employed for the resolution of SSR amplified products. Physiologically mature leaf was found more suitable for getting quality DNA in mulberry.
Highlights
Extraction of quality DNA is a major challenge for molecular biologists dealing with tree plants
We report a simple, rapid and efficient method yielding appreciable levels of high quality DNA suitable for molecular studies including restriction digestion and Polymerase Chain Reaction (PCR) amplifications
Rotundiloba, M. serrata, M. laevigata and M. nigra grown in the germplasm of Central Sericultural Germplasm Resources Center (CSGRC), Hosur, Tamil Nadu, India were selected for the study
Summary
Extraction of quality DNA is a major challenge for molecular biologists dealing with tree plants. Higher contents of polyphenolic compounds, resins, latex, and polysaccharides, tannins present in the cell as secondary metabolites usually co precipitate with DNA and interfere with the activity of the DNA polymerase enzyme (Pandey et al, 1996). H. Jingade Anuradha Scientist C, Central Sericultural Germplasm Resources Centre, Central Silk Board, Thally Road, Hosur-635 109, Tamil Nadu, India. Jingade Anuradha Scientist C, Central Sericultural Germplasm Resources Centre, Central Silk Board, Thally Road, Hosur-635 109, Tamil Nadu, India Usage of these commercial kits for routine extraction is economically difficult for large-scale genomic applications. We report a simple, rapid and efficient method yielding appreciable levels of high quality DNA suitable for molecular studies including restriction digestion and PCR amplifications. A protocol of silver staining the Polyacrylamide Gel Electrophoresis (PAGE) gel for microsatellite analysis in mulberry is established in the present study
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