A Simple Procedure for Extraction and Purification DNA in Chinese fir

A simple, quick and reliable procedure was developed for the extraction and purification of DNA from the leaves of Chinese fir.Specifically designed to overcome the trouble caused by the very high concentration of pigments,polysaccharides and other contaminants in the leaves.In this procedure,the application of extraction buffer with high concentration of mercaptoethanol (2%)was proved to be successful in removing pigments,ethanol precipitation of resuspended nucleic acids from 3 mol/L NaAC and revealed to be powerful in removing polysaccharides,and PVP-360 was used to remove polyphenols.The resulted DNA with this method was of high molecular weight (more than 50 kb) and was completely digestible with restriction endonucleases and amplifiable in PCR,they are well suited to RAPD,RFLP analysis and other molecular marker systems.This procedure can also be used to extract DNA from other gymnosperms,for example:Taiwania fiousiana,Metasequoia glyptostroboides,Cryptomeria fortunei etc.