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Harfe BD, & Fire A (1997). New RNA in situ fixative. Worm Breeder's Gazette, g14, p2. Unpublished information; cite only with author permission.
Recently, we learned of a new, cheap fixative that works very well on sea urchin embryos [S.T.F. Molecular Biology Grade, Streck Laboratories (1-800-228-6090)]. We have begun to use this fix instead of the formaldehyde fix commonly used in worm RNA in Situ's. We are following the procedure of Seydoux and Fire, except for the substitution of an overnight incubation at room temperature in S.T.F fixative instead of the fixation step from the Seydoux procedure. With a ceh-27 probe, we see a hybridization signal about three times stronger using the Streck fix than with the standard formaldehyde fix. We have found that the S.T.F. fix can be stored at room temperature and reused (at least six times). General in situ procedure: G. Seydoux and A. Fire. Whole-Mount in Situ Hybridization for the Detection of RNA in Caenorhabditis elegans Embryos. In "Caenorhabditis elegans, Modern Biological Analysis of an Organism" (edited by H. Epstein and D. Shakes) 1995. We thank C. Kirchhamer and G. Seydoux.
Affiliation:
- The Johns Hopkins University Biology Graduate Program, The Carnegie Institution of Washington The Carnegie Institution of Washington