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Simple method to prepare DNA templates from a slice of peanut cotyledonary tissue for Polymerase Chain Reaction Shu Tao Yu# Chuan Tang Wang*# Shan Lin Yu# Xiu Zhen Wang Yue Yi Tang Dian Xu Chen Jian Cheng Zhang *Corresponding author #All of the three authors contributed equally Financial support: Modern Agro-Industry Technology Research System (MATRS) Peanut Program, Ministry of Agriculture, China; China Natural Science Foundation (Grant No. 30300224), 863 New and High Technology Project (Grant No. 2006AA10A114), New and High Technology Innovation Foundation of Shandong Academy of Agricultural Sciences (Grant No. 2006 YCX013). Young Scientists Foundation of Shandong Academy of Agricultural Sciences (Grant No. 2007YQN007), Shandong Natural Science Foundation (Grant No. Y2008D11), and Shandong Key Projects for Science and Technology (Grant No. 2009GG10009008). Keywords: cotyledonary tissue, DNA extraction, groundnut, PCR, peanut.
An efficient DNA extraction method was developed for peanut seed, where only 3-5 mg cotyledonary tissue was enough for more than 50 PCR reactions with a reaction volume of 15 μl. Both low copy number and high copy number DNA sequences were successfully amplified. Processing one seed sample only took about half an hour. Sampling had no significant effects on germination and development. The DNA extraction method makes it possible to identify transformants and conduct molecular marker studies prior to sowing, and thus may greatly hasten research progress. |