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Abstract [Objective] The aim was to seek for a rapid DNA minipreparation method from wheat leaf. [Method] The total DNA of wheat leaf was extracted using CTAB, SDS and boiling water, separately, with some modifications. Integrity and purity of nucleic acids were detected through agarose gel electrophoresis, ultraviolet absorption and PCR. [Result] The DNA extracted by the modified CTAB method had high quality and purity, and was not degraded. Two hundreds of DNA samples could be extracted each workday by per capita using this method; and the PCR detection of wheat transgenic plants showed that amplified bands of target gene were clear, without false-positive, and the test results were satisfactory. The DNA purity and concentration extracted by modified SDS method were not as good as that extracted by modified CTAB method, but it also met the DNA requirements of major molecular research. The DNA quantity extracted by modified boiling method was small and there were a lot of impurities in it, PCR detection of this DNA showed no amplified band. [Conclusion] Modified CTAB method is a simple and rapid method for DNA minipreparation from wheat leaf, and was suitable for PCR amplification and other molecular biology researches.
Key words Wheat; DNA extraction; Modified CTAB method
(ProQuest: ... denotes non-USASCII text omitted.)
DNA extraction from plant is the most basic procedure in molecular biology. The DNA quality is very critical, because it directly affects all the following steps[1-2]. So far, various methods for extracting DNA from plants have been reported, such as SDS method[3] and CTAB method[4]. By these methods, the isolated DNA has high quality and purity, and can meet the requirements of PCR analysis. But these methods involve tedious procedures, which reduce the efficiency of molecular marker detection, and thereby influence the popularization and promotion of molecular markers[5-7]. In recent years, a variety of simple methods for DNA extraction have been developed, such as alkali treatment[8], boiling water method[9], SDS method[10] and grinding with TE[11]. Most of these methods are rapid and low-cost, using no liquid nitrogen [12] and phenol113-141. But, the DNA extracted by these methods usually has poor quality and can not be amplified by PCR [15"161. These methods can not meet the requirement for preparing a large amount of...