Analysis of methylation and distribution of CpG sequences on human active and inactive X chromosomes by in situ nick translation

Chromosoma ◽  
1990 ◽  
Vol 99 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Giorgio Prantera ◽  
Marina Ferraro
Keyword(s):  
Nick Translation ◽  
X Chromosomes ◽  
In Situ Nick Translation

In situ nick-translation distinguishes between active and inactive X chromosomes

Nature ◽  
1983 ◽  
Vol 304 (5921) ◽  
pp. 88-90 ◽  
Author(s):  
Bat-Sheva Kerem ◽  
Ruth Goitein ◽  
Carmelit Richler ◽  
Menashe Marcus ◽  
Howard Cedar
Keyword(s):  
Nick Translation ◽  
X Chromosomes ◽  
In Situ Nick Translation

The distribution of genes on human chromosomes as studied by in situ nick translation

Genome ◽  
1992 ◽  
Vol 35 (5) ◽  
pp. 890-894 ◽  
Author(s):  
J. de la Torre ◽  
A. T. Sumner ◽  
J. Gosalvez ◽  
L. Stuppia
Keyword(s):  
Cpg Islands ◽  
Dnase I ◽  
Human Chromosomes ◽  
Nick Translation ◽  
X Chromosomes ◽  
Dnase I Hypersensitivity ◽  
In Situ Nick Translation ◽  
Inactive X Chromosome ◽  
Active Genes

We have studied the distribution of potentially active genes on human chromosomes, using two methods: DNAse I hypersensitivity and restriction enzyme – nick translation with enzymes sensitive to methylation of CpG doublets. DNAse hypersensitivity is known to be associated with potentially active genes, and, when the reaction is detected by "in situ" nick translation, produces an R-banding pattern. Digestion of chromosomes with HpaII or CfoI, both of which should preferentially cut unmethylated sequences in the CpG islands associated with the majority of genes, also produces R-banding patterns. Deviations are attributable to overdigestion of the chromosomes, leading to extraction of DNA and loss of the specific sites that were to be detected. Contrary to the results of a number of previous workers, we have failed to demonstrate any differences between the DNAse I hypersensitivity or the degree of methylation of the active and inactive X chromosomes in metaphases from females.Key words: human chromosomes, gene distribution, DNAse I hypersensitivity, in situ nick translation, R-banding, CpG islands, DNA methylation, inactive X chromosome.

scholarly journals Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections.

1993 ◽  
Vol 41 (7) ◽  
pp. 1023-1030 ◽  
Author(s):  
R Gold ◽  
M Schmied ◽  
G Rothe ◽  
H Zischler ◽  
H Breitschopf ◽  
...  
Keyword(s):  
Cell Death ◽  
Dna Fragmentation ◽  
Specific Cell ◽  
Nick Translation ◽  
Proliferating Cells ◽  
Cell Surface Antigens ◽  
Simultaneous Application ◽  
Tissue Sections ◽  
In Situ Nick Translation

Since DNA fragmentation is a key feature of programmed cell death (PCD) and also occurs in certain stages of necrosis, we have adapted the methodology of in situ nick-translation (ISNT) to detect DNA fragmentation on a single-cell level. We first established the technique for cell preparations. Apoptosis was induced by gamma-irradiation on freshly isolated rat thymocytes. After fixation procedures, ISNT was performed by overnight incubation either with fluorescein-12-dUTP or with digoxigenin-labeled 11-dUTP and DNA polymerase I. The enzymatic incorporation of labeled nucleotides at sites of DNA fragmentation was detected by flow cytometry either directly or indirectly with fluorescein-conjugated anti-digoxigenin. The quantitative results demonstrated close correlation with morphological essays for apoptosis, DNA gel electrophoresis, and ISNT. Proliferating cells determined by bromodeoxyuridine immunofluorescence were not labeled by ISNT. Immunocytochemistry for cell surface antigens in combination with ISNT allowed the identification of specific cell types undergoing PCD. Furthermore, the simultaneous application of photolabeling techniques with ethidium monoazide and ISNT led to the identification of DNA fragmentation in cells with still intact membranes. Extending ISNT to tissue sections of paraformaldehyde-fixed, paraffin-embedded material reliably revealed labeling of cells with typical morphological features of apoptosis. However, this technique was not useful in detecting early stages of necrotic cell death.

Microprocedure for in situ nick translation of chromosomes

1992 ◽  
Vol 62 (2) ◽  
pp. 150-153
Author(s):  
Li Zhengang ◽  
Howard L. Hosick ◽  
Kang Fan
Keyword(s):  
Nick Translation ◽  
In Situ Nick Translation

DNA single-strand breaks in postischemic gerbil brain detected by in situ nick translation procedure

1995 ◽  
Vol 200 (2) ◽  
pp. 129-132 ◽  
Author(s):  
Muneshige Tobita ◽  
Isao Nagano ◽  
Shozo Nakamura ◽  
Yasuto Itoyama ◽  
Kyuya Kogure
Keyword(s):  
Single Strand ◽  
Nick Translation ◽  
Strand Breaks ◽  
In Situ Nick Translation ◽  
Gerbil Brain ◽  
Single Strand Breaks ◽  
Translation Procedure

In situ nick translation of meiotic chromosomes to demonstrate homologous heterochromatin heterogeneity

Genome ◽  
1993 ◽  
Vol 36 (2) ◽  
pp. 268-270 ◽  
Author(s):  
J. de la Torre ◽  
C. López-Fernández ◽  
P. Herrero ◽  
J. Gosálvez
Keyword(s):  
Tandem Duplication ◽  
Dna Recombination ◽  
Nick Translation ◽  
Meiotic Chromosomes ◽  
In Situ Nick Translation ◽  
Living Species ◽  
Longitudinal Differentiation ◽  
Translation Procedure ◽  
Restriction Endonuclease Cleavage

The in situ nick translation procedure performed on fixed meiotic chromosomes partially cleaved with restriction endonucleases shows a different staining of homologous heterochromatic regions, which could be explained through a differential restriction endonuclease cleavage. Mutations occurring before massive tandem duplication and involving those DNA motifs that produce these heterochromatic blocks, together with the absence of DNA recombination that characterizes these particular regions, could explain the observed results. This method for chromosome labelling is most useful to demonstrate a certain level of heterochromatin heterogeneity that is present in the genome of living species but remained cryptic to other techniques that are also able to induce longitudinal differentiation of the chromosomes.Key words: cytogenetics, nick translation, meiosis, heterochromatin.

A new banding pattern of human chromosomes by in situ nick translation using ECO RI and biotin-dUTP

2008 ◽  
Vol 28 (2) ◽  
pp. 173-176 ◽  
Author(s):  
Jörn Bullerdiek ◽  
Jürgen Dittmer ◽  
Angelika Faehre ◽  
Sabine Bartnitzke ◽  
Volker Kasche ◽  
...  
Keyword(s):  
Banding Pattern ◽  
Human Chromosomes ◽  
Nick Translation ◽  
In Situ Nick Translation ◽  
Eco Ri
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