Restriction enzyme banding of mouse metaphase chromosomes

Chromosoma ◽  
1984 ◽  
Vol 90 (2) ◽  
pp. 128-132 ◽  
Author(s):  
Margot Kaelbling ◽  
Dorothy A. Miller ◽  
Orlando J. Miller
Keyword(s):  
Restriction Enzyme ◽  
Metaphase Chromosomes ◽  
Restriction Enzyme Banding

Comparison of methods for the detection of in situ restriction enzyme – nick translation using fluorochromes and confocal microscopy

Genome ◽  
1995 ◽  
Vol 38 (5) ◽  
pp. 1032-1036 ◽  
Author(s):  
L. Stuppia ◽  
C. Cinti ◽  
S. Santi ◽  
R. Peila ◽  
N. M. Maraldi ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Restriction Enzyme ◽  
Restriction Enzymes ◽  
Chromosome Morphology ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Direct Labelling ◽  
Series Of Experiments ◽  
The Cost

A series of experiments was carried out to determine the most efficient methods for detecting incorporated nucleotides in the "in situ" restriction enzyme – nick translation technique. Different methods were tested on fixed human metaphase chromosomes using confocal microscopy for the demonstration of the patterns produced. Of the various techniques tested, that using DIG-dUTP in conjunction with FITC-labelled anti-DIG appears to show the greatest sensitivity and specificity. The use of biotinylated nucleotides with FITC-avidin gives rather less sensitivity, while direct labelling with fluorescein-dUTP produces results more rapidly with better chromosome morphology but at the cost of reduced sensitivity. Resorufin-labelled dUTP was unusable, because of the low level of fluorescence and its very rapid fading. The successful fluorescence methods are more sensitive and faster than using horseradish peroxidase or alkaline phosphatase for detection.Key words: restriction enzymes, nick translation, chromosomes, fluorochromes, confocal microscopy.

The effect of restriction enzyme digestion of human metaphase chromosomes on C-band variants of chromosomes 1 and 9

Genome ◽  
1988 ◽  
Vol 30 (5) ◽  
pp. 652-655 ◽  
Author(s):  
Ute Hedemann ◽  
M. Schürmann ◽  
E. Schwinger
Keyword(s):  
Restriction Enzyme ◽  
Restriction Enzymes ◽  
Enzyme Digestion ◽  
Restriction Endonucleases ◽  
Restriction Enzyme Digestion ◽  
Human Chromosomes ◽  
Metaphase Chromosomes ◽  
Homologous Chromosomes

Human metaphase chromosomes, fixed on slides, have been treated with 8 different restriction endonucleases and 29 combinations of 2 restriction enzymes prior to staining with Giemsa. The endonucleases AluI and DdeI and the combinations AluI + DdeI, AluI + HaeIII, AluI + HinfI, and AluI + MboI have then been used to digest metaphase chromosomes of nine individuals with C-band variants of chromosomes 1 or 9, obtained by the CBG technique. The restriction enzyme resistant chromatin of the paracentromeric regions of chromosomes 1 and 9 has been measured and compared with the corresponding CBG-bands. The size of the enzyme resistant chromatin regions depend upon the type of enzyme(s) used. Treatment with AluI + MboI was the only digestion that acted differently on different chromosome pairs. However, within one pair of homologous chromosomes, all digestions revealed the same variations as conventional C-banding.Key words: C-band variants, heterochromatin, human chromosomes, restriction endonucleases.

scholarly journals Restriction enzyme digestion chromosome banding in Crassostrea and Ostrea species: comparative karyological analysis within Ostreidae

Genome ◽  
2004 ◽  
Vol 47 (5) ◽  
pp. 781-788 ◽  
Author(s):  
A Leitão ◽  
R Chaves ◽  
S Santos ◽  
H Guedes-Pinto ◽  
P Boudry
Keyword(s):  
Restriction Enzyme ◽  
Chromosome Banding ◽  
Genome Mapping ◽  
Genetic Research ◽  
Restriction Enzymes ◽  
Restriction Enzyme Digestion ◽  
Banding Patterns ◽  
Crassostrea Angulata ◽  
Restriction Enzyme Banding

Reliable banding techniques are a major necessity for genetic research in oysters. In this study, we carried out the cytogenetic characterization of four oyster species (family Ostreidae) using restriction endonuclease treatments. Chromosomes were treated with three different restriction enzymes, stained with Giemsa, and examined for banding patterns. The following species were studied: Crassostrea gigas (2n = 20; total number of bands with ApaI, 74; HaeIII, 61; PstI, 76), Crassostrea angulata (2n = 20; ApaI, 62; HaeIII, 61; PstI, 55) (subfamily Crassostreinae), Ostrea edulis (2n = 20; ApaI, 82; HaeIII, 59; PstI, 66), and Ostrea conchaphila (2n = 20; ApaI, 68; HaeIII, 62; PstI, 69) (subfamily Ostreinae). Treatment of samples with ApaI, HaeIII, and PstI produced specific banding patterns, which demonstrates the potential of these enzymes for chromosome banding in oysters. This is of special interest, since it has been recently shown in mammalian chromosomes that restriction enzyme banding is compatible with fluorescence in situ hybridization. This study therefore provides a fundamental step in genome mapping of oysters, since chromosome banding with restriction enzymes facilitates physical gene mapping in these important aquaculture species. The analysis of the banded karyotypes revealed a greater similarity within the genera of Crassostrea and Ostrea than between them.Key words: Ostreidae, Crassostrea, Ostrea, chromosome banding, in situ restriction enzyme banding.

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