整体荧光原位杂交用于检测水稻特异DNA程序的研究

STUDY ON THE PROCEDURE OF WHOLE MOUNT FLUORESCENCE IN SITU HYBRIDIZATION FOR DETECTING SPECIFIC DNA IN RICE

摘要: 文章报道了整体荧光原位杂交应用于检测水稻特异ＤＮＡ的实验程序。利用该实验方法，以经地高辛（ＤＩＧ）标记的ｒＤＮＡ作为探针，成功地在水稻根尖和花药内部组织检测与探针互补的片段。对于实验方法，结果表明，整体荧光原位杂交的成功与否受到多种因素的影响，包括组织的种类、固定剂的种类以及固定后的处理方法（主要是酶解与否）等。检测根尖ＤＮＡ可以采用φ（福尔马林）＝１％固定加上酶解；检测花药（花粉发育处于单核期）

Abstract: A procedure of whole mount flurescence in situ hybridization (WFISH) for detecting specific DNA in rice was reported. The techique was demonstrated on root and anther of rice with rDNA as probe, labelled by DIG. It was found that the hybridization efficiency depended on the methods further used after fixation, types of tissue and of fixative. Fixation in 1% formalin plus enzyme digestion could yield excellent results of DNA:DNA in root tips. Fixation in 1% formalin (no need enzyme digestion) yielded the excellent results with respect to morphology and hybridization efficiency in rice anther.