scholarly journals BASIC FUCHSIN AND THE FEULGEN REACTION: SIGNIFICANCE OF THE DYE FOR CYTOPHOTOMETRIC DETERMINATION OF DEOXYRIBONUCLEIC ACID IN CELL NUCLEI

1963 ◽  
Vol 11 (3) ◽  
pp. 401-408 ◽  
Author(s):  
Z. LODIN ◽  
J. MÜLLER ◽  
J. PILNÝ ◽  
J. HARTMAN
1961 ◽  
Vol 9 (3) ◽  
pp. 619-626 ◽  
Author(s):  
Jan-Erik Edström ◽  
Jerzy Kawiak

A method for the quantitative determination of DNA in the 50 to 500 µµg. range is presented. Cells or cell nuclei are isolated individually from fixed tissue by means of micromanipulation. The tissue units in question are extracted in an oil chamber with deoxyribonuclease solution. The extracts are evaporated to dryness and redissolved to lens-shaped drops, the DNA contents of which are determined by a photographic-photometric procedure in ultraviolet light. Determinations on calf thymocytes and rat spermatids show a relatively good agreement with biochemical data. The present method tends, however, to give some. what higher values than those reported earlier. The coefficient of variation for analytical values from test material is about ± 10 per cent. The method has been applied to cells from the axolotl, adults as well as tadpoles. Germ cells (spermatids and spermatocytes) do not show any evidence of a biological variation in DNA content. Cells from proliferating tissues give an increased spread of the DNA values. It could be shown, for epithelial cells, that there are at least two factors determining the DNA content of these cells. One is the fact that the cells are investigated at different phases of the mitotic cycle; the other is the fact that the DNA synthesis cycle occupies different ranges for different cells.


1964 ◽  
Vol 12 (10) ◽  
pp. 758-767 ◽  
Author(s):  
M. J. HARDONK ◽  
P. VAN DUIJN

The absorption spectra of Feulgen-stained deoxyribonucleic acid (DNA)-containing membranes have been studied under various conditions. The absorption curve of the Feulgen-stained DNA-cellulose differs from those of the other model systems and is comparable to that of a solution of apurinic acid stained by the Schiff reaction. The spectral shift caused by the reaction of formaldehyde and SO2 with the stained films was used as a criterion for the degree of substitution of the bound pararosaniline. The spectrum of DNA-cellulose, unlike the spectra of the other models, did not show any change under the action of these reagents. This indicates that the three amino groups of pararosaniline are all bound to the DNA in a stained DNA-cellulose film. When the mobility of the apurinic acid chain is restricted, the formation of the trisubstituted product is apparently hindered. Such a situation may be present in DNA-containing polyacrylamide films containing proteins and also in cell nuclei. From a study of the literature it is concluded that the shoulder in the pararosaniline spectrum is most probably caused by a configurational isomer of the dye as indicated by Lewis et al. (16) for crystal violet. To explain the abnormal spectrum of the stained DNA-cellulose, it is proposed that in this case pararosaniline is bound with the three amino groups to three neighbouring deoxyribose groups in the hydrolyzed DNA. Studies with Stuart models of the molecules involved showed that in this case one conformation isomer of pararosaniline can be constructed more easily than the other. This situation may explain the spectrum of the stained DNA-cellulose. The other spectral results could also be interpreted on basis of this hypothesis.


2011 ◽  
Vol 3 (1) ◽  
Author(s):  
Lies Indah Sutiknowati

There is an information how to identify hydrocarbon degrading bacteria for bioremediation of marine oil spill. We have Bioremediation treatment for degradation of oil spill on Pari island and need two kind of experiment there are tanks experiment (sampling 0 to 90 days) and semi enclosed system (sampling 0 to 150 days). Biostimulation with nutrients (N and P) was done to analyze biodegradation of hydrocarbon compounds. Experiment design using fertilizer Super IB and Linstar will stimulate bacteria can degrade oil, n-alkane, and alkane as poly aromatic hydrocarbon. The bacteria communities were monitored and analyzed by Denaturing Gradient Gel Electrophoresis (DGGE) and Clone Library; oil chemistry was analyzed by Gas Chromatography Mass Spectrometry (GCMS). DNA (deoxyribonucleic acid) was extracted from colonies of bacteria and sequence determination of the 16S rDNA was amplified by primers U515f and U1492r. Strains had been sequence and had similarity about 90-99% to their closest taxa by homology Blast search and few of them suspected as new species. The results showed that fertilizers gave a significant effect on alkane, PAH and oil degradation in tanks experiment but not in the field test. Dominant of the specific bacteria on this experiment were Alcanivorax, Marinobacter and Prosthecochloris. Keywords: Bioremediation, Biostimulation, DGGE, PAH, Pari Island


1963 ◽  
Vol 238 (3) ◽  
pp. 1068-1072
Author(s):  
Peter Schmid ◽  
Charlotte Schmid ◽  
Donald C. Brodie

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.


1969 ◽  
Vol 47 (9) ◽  
pp. 1367-1375 ◽  
Author(s):  
T. A. Steeves ◽  
M. Anne Hicks ◽  
J. M. Naylor ◽  
Patricia Rennie

The vegetative shoot apex of Helianthus annuus contains a central zone in which the cell nuclei are relatively large and stain faintly in the Feulgen reaction. Excised apices in the vegetative state were supplied with thymidine-H3 through their sterile, liquid culture medium. Autoradiography after 24 or 48 hours of feeding revealed no significant incorporation of the labeled precursor into central zone nuclei, but extensive incorporation in peripheral regions of the apex. It is concluded that during vegetative growth deoxyribonucleic acid (DNA) synthesis and mitosis are arrested in the central zone or reduced to an extremely slow rate. Microspectrophotometry, however, indicates that the central zone nuclei are not held at the 2C level. With the onset of flowering, cytological zonation disappears in the apex and the incorporation of thymidine-H3 is uniformly heavy throughout the apical region.


1989 ◽  
Vol 160 (3) ◽  
pp. 734-736 ◽  
Author(s):  
Chihiro Azuma ◽  
Shoji Kamiura ◽  
Toshikatsu Nobunaga ◽  
Takao Negoro ◽  
Fumitaka Saji ◽  
...  

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