By Claudio Nicolini
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Additional resources for Chromatin Structure and Function: Molecular and Cellular Biophysical Methods
LANGLOIS: Do metaphase chromosomes have a different structure at the nucleosome level than interphase chromatin? DR. BONNER: I do not know; perhaps people working at the E. M. level will be able to tell us. DISCUSSION 27 DR. NICOLINI: I am disturbed by the lack of interest and precision in defining the object of your analysis and how much it could be different from the original object "in situ". Without such an effort it is difficult to communicate. In studying higher levels of organization such standardization is extremely important.
Ve gene expression can be seen in in vivo made RNA has been re-examined from the point of view of messenger sequences. This has been R. S. GILMOUR done by hybridizing RNA to the unique DNA sequences isolated from the total genomic DNA or to cDNA made to specific mUNA. Under these hybridization conditions the concentrations of the unique sequences is effectively concentrated and hence can undergo hYbridization. For example, transcripts derived from unique DNA sequences were analysed by hybridising isolated unique DNA to a large excess of total cellular RNA or in some cases nuclear UNA.
This does not, of course, apply to Xerox copies of micrographs, transpariencies projected in an illuminated room or slides viewed at a great distance. Biological specimens are composed of small atoms which have little intrinsic electron density, therefore most contrast is obtained by the addition of electron dense molecules (stains). Positive staining is achieved by incubating the specimen with the stain followed by washing the specimen in order to remove all unbound stain. This method requires a high affinity between stain and specimen.