scholarly journals Direct Imaging of Northern Blots on an Optical Scanner Using Ethidium Bromide

BioTechniques ◽  
2000 ◽  
Vol 28 (5) ◽  
pp. 864-870 ◽  
Author(s):  
Roberta L. Eykholt ◽  
Murray D. Mitchell ◽  
Keith W. Marvin
Author(s):  
K. S. McCarty ◽  
R. F. Weave ◽  
L. Kemper ◽  
F. S. Vogel

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.


Author(s):  
E. Keyhani

The mutagenic effect of ethidium bromide on the mitochondrial DNA is well established. Using thin section electron microscopy, it was shown that when yeast cells were grown in the presence of ethidium bromide, besides alterations in the mitochondria, the plasma membrane also showed alterations consisting of 75 to 110 nm-deep pits. Furthermore, ethidium bromide induced an increase in the length and number of endoplasmic reticulum and in the number of intracytoplasmic vesicles.Freeze-fracture, by splitting the hydrophobic region of the membrane, allows the visualization of the surface view of the membrane, and consequently, any alteration induced by ethidium bromide on the membrane can be better examined by this method than by the thin section method.Yeast cells, Candida utilis. were grown in the presence of 35 μM ethidium bromide. Cells were harvested and freeze-fractured according to the procedure previously described.


2012 ◽  
Vol 132 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Satoshi Maruyama ◽  
Muneki Nakada ◽  
Makoto Mita ◽  
Takuya Takahashi ◽  
Hiroyuki Fujita ◽  
...  

2012 ◽  
Vol 11 (9) ◽  
pp. 1721-1724
Author(s):  
Roxana Vatavu ◽  
Constantin Leonte ◽  
Teodor Robu. ◽  
Catalina Slabu-Pascal

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