| Autor: |
Goto, Toshiyuki, Kohno, Takehiro, Nakano, Takashi, Fujioka, Yoshihiko, Morita, Chizuko, Sano, Kouichi |
| Zdroj: |
Journal of Electron Microscopy; Jan2001, Vol. 50 Issue 1, p73-76, 4p |
| Abstrakt: |
Electron microscopic in situ hybridization (EM‐ISH)is a useful method in determining the localization of a specific nucleic acidat the ultrastructural level. Since the EM‐ISH protocol includes manysteps, no standard protocol for EM‐ISH is available yet. In this study,we optimized quantitatively the critical conditions with respect to embeddingresin, nucleic acid labeling and hybridization reaction time, by using adenovirus‐infectedcells as the indicator cells. The optimal detection of an adenovirus‐specificnucleic acid was obtained by overnight hybridization reaction on sectionsembedded in Lowicryl K4M resin. Random‐primed‐labeled probesimproved the reactivity. At least 60% of virus particles in paracrystallinearrays was found to contain viral DNA. These arrays in adenovirus‐infectedcells are useful in evaluating quantitatively the efficiency of protocolsof EM‐ISH. [ABSTRACT FROM PUBLISHER] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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