In Situ Hybridization on Plant Metaphase Chromosomes
In very recent years, the methods of molecular biology are being applied in plant cytogenetics. In situ hybridization is one of these methods. It is the hybridization of a nucleic acid probe (RNA or DNA) to nucleic acids within cytological preparations. It is obvious that gene localization on metaphase chromosomes requires a highly sensitive in situ hybridization method that makes chromosomal DNA accessible to the probe. The accessibility of the target DNA may be facilitated in plant cells by: (1) removing the cell wall, i.e., protoplasting the plant cells, (2) increasing the sensitivity and the efficiency of hybridization, and (3) denaturing very efficiently both probe and chromosomal DNA.
KeywordsBoiling Cellulase Xylene Triphosphate Colchicine
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