By John M. Walker
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Extra info for New Nucleic Acid Techniques
USA 78,2253-2257. Fischer, S. G. (1983) Peptide mapping in gels. M&h. Enzymol. 100,424430. , and Tucker, G. A. (1985) The appearance of polygalacturonase mRNA in tomatoes: One of a series of changes in gene expression during development and ripening. Planta 163,263-271. Jackson, R. C. and Blobel, G. (1977) Post-translational cleavage of presecretory proteins with an extract of rough microsomes from dog pancreas containing signal peptidase activity. Nafl. Acad. Sci. USA 74,5598-5602. Noyes, B. E.
100,424430. , and Tucker, G. A. (1985) The appearance of polygalacturonase mRNA in tomatoes: One of a series of changes in gene expression during development and ripening. Planta 163,263-271. Jackson, R. C. and Blobel, G. (1977) Post-translational cleavage of presecretory proteins with an extract of rough microsomes from dog pancreas containing signal peptidase activity. Nafl. Acad. Sci. USA 74,5598-5602. Noyes, B. E. and Stark, G. R. (1975) Nucleic acid hybridisation using DNA covalently coupled to cellulose.
Cellular or cytoplasmic) RNA, and this has a number of advantages compared to using poly(A)+ RNA. The large proportion of ribosomal RNA in the sample acts as a carrier, minimizing nonspecific binding of mRNA to the filter and reducing the effect of ribonuclease contamination. Furthermore, the time required to isolate sufficient RNA for this procedure may itself be an important consideration. The order of addition of hybridization buffer constituents to total RNA is critical in that a precipitate may form if the 4M NaCl is added before the other components.