A convenient and efficient procedure for isolation of rice HMW DNA bad been developed.Etiolated seedings were ground into the fine powder with a pestle and mortar under liquid nitrogen,then extracted with a protective medium,Filtrate was layered on discontinuous sucrose gradient,then the purified nuclei were embeded in agarose gel.After proteinase digestion,the HMW DNA was released. Pulsed-field gel electrophoresis showed that the sample size ranges from 200 kb to 3 Mb and in the majority of 2.2 Mb. Rice YAC library was constructed successfully, using these easily digestable DNA preparation.
Wang Chunxin, Xie Yiwu, Liu Liangshi. Isolation of Megabase DNA from Rice Nuclei[J]. Progress in Biochemistry and Biophysics,1995,22(4):357-361
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