杉木、杨树激光显微切割体系的建立及RNA质量的控制-遗传学专业论文.docxVIP

杉木、杨树激光显微切割体系的建立及RNA质量的控制-遗传学专业论文.docx

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The The Laser Microdissection system of Chinese fir and Popuplus cambium and RNA quality control Abstract Laser mierodissection(LMD)is a technique which can isolate specific tissues,cell types and eveu organeHes from sectioned biological specimen under the microscope in a manlier conducive to the extraction of RNA.DNA or protein.Pioneering research has demonstrated the potential of LMD as a powerful tool for investigating molecular and biochemical events that occur in discrete cells ofcomplex,heterogeneous tissues。with the characteristic offast,precise and easy-operation.LMD,which is an established technique in many areas of biology,has now been successfully adapted for use wi血plant tissues. Woody plants have the ability to produce secondary tissue(secondary xylem and phloem)from a medstematic tissue called vascular cambium,responsible for the radial growth of a tree.The research ofexpression and control ofgene during the wood formation by molecular and cytological is an important scientific issue for scientists.However,the knowledge of cambium is limited.because it is hard to separate the cambium ceIls from other surrounding cells by traditional methods.Selecting the cambium cells precisely is a key point for the research of cambium cells.Since 1990s,the development of Laser Microdissection facilitated the collection of special materials. In the first step of research,we established a suitable cryo—sectioning method for stem organs of Chinese fir and Populus.With studying on the technical parameters of fixation and pretreatment Of tissue.cultured material.a suitable procedure consisted of several steps is considered the best protocol as follows.The samples were first fixed in Canoy’s fixation for 4 to 6 hours,and then saturated in 10%sucrose PBS solution for 6 to 12 hours.After embedded they were frozen in fiquid nitrogen for 50 to 90 seconds and cryo·sectioned.The procedure of dehydration after cryo—sectioning improved the quality of section to a certain extent.T

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