In Situ Hybridisation: Application to Developmental Biology by N. Harris (editor), D. G. Wilkinson (editor)

By N. Harris (editor), D. G. Wilkinson (editor)

Advances in our knowing of organic mechanisms have often been linked to the improvement of recent innovations. In situ hybridization is a vintage case of simply such an boost. The strategy successfully combines histochemistry with molecular biology and permits the fast research of the distribution of RNA, or DNA, within the tissues. the data received from this has brought on whatever of a revolution in our realizing of developmental biology, because a primary point of improvement is the spatial and temporal expression of genes. furthermore the approach has stumbled on program within the box of medication, the place it truly is giving new insights into the functioning of fit tissues and the analysis and research of ailments. This ebook brings jointly contributions from leaders within the software of this method and should support the reader evaluate and be guided during the a variety of recommendations and adaptations of the method.

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Extra info for In Situ Hybridisation: Application to Developmental Biology and Medicine (Society for Experimental Biology Seminar Series (No. 40))

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Landegant, J. E. (1987). Non-radioactive in situ hybridization: a comparison of several immunocytochemical detection systems using reflection contrast and electron microscopy. Histochemistry, 86(6), 609-15. Crum, C. , Friedman, D & Silverstein, S. J. (1988). A comparison of biotin and isotope-labeled ribonucleic acid probes for in situ detection of HPV-16 ribonucleic acid in genital precancers. Laboratory Investigation, 58 (3), 354-9. , Krause, J. , Polak, J. M. & Legon, S. (1988). Oligoriboprobes: tools for in situ hybridization.

Fluorescence in situ hybridisation 35 For in situ DNA detection, target DNA denaturation has to be performed. There seems to be general consensus that heat denaturation with or without probe already present yields good results. Other significant points are the use of formamide in hybridisation solutions to reduce optimal hybridisation temperatures, and the fragmentation of probe molecules so as to promote probe penetration. In this respect synthetic oligonucleotides can be very useful. Their efficient in situ hybridisation, which is probably due to their good penetration properties, may in fact compensate to a large extent for the smaller target they cover (Dirks et al, in preparation).

Gait, J. C. (1986). Efficient methods for attaching non-radioactive labels to the 5' end of synthetic oligodeoxyribonucleotides. Nucleic Acids Research, 14, 6227-45. Albertson, D. G. (1985). Mapping muscle protein genes by in situ hybridization using biotin-labelled probes. The EMBO Journal, 4, 2493-8. Ambros, P. , Matzke, M. A. & Matzke, A. J. M. (1986). Detection of a 17 kb unique (T-DNA) in plant chromosomes by in situ hybridization. Chromosoma, 94, 11-16. Appels, R. (1989). Three dimensional arrangement of chromatin and chromosomes: old concepts and new techniques.

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