Nick-Translation Reaction Buffer (10×)

doi:10.1101/pdb.rec104547

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122885 ◽

1992 ◽

Vol 50 (1) ◽

pp. 496-497

Author(s):

Barbara Trask ◽

Susan Allen ◽

Anne Bergmann ◽

Mari Christensen ◽

Anne Fertitta ◽

...

Keyword(s):

In Situ Hybridization ◽

Dna Sequence ◽

Dna Sequences ◽

Dual Band ◽

Nick Translation ◽

Metaphase Chromosomes ◽

Band Pass ◽

Texas Red ◽

Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

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A Modified Nick Translation Method Used with FISH that Produces Reliable Results with Archival Tissue Sections

Molecular Biotechnology ◽

10.1385/mb:20:3:257 ◽

2002 ◽

Vol 20 (3) ◽

pp. 257-260 ◽

Cited By ~ 1

Author(s):

Amanda D Watters ◽

Michael W Stacey ◽

John M S Bartlett

Keyword(s):

Translation Method ◽

Nick Translation ◽

Archival Tissue ◽

Tissue Sections

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Molecular characterization of Vibrio anguillarum biotype 2

Canadian Journal of Microbiology ◽

10.1139/m81-158 ◽

1981 ◽

Vol 27 (10) ◽

pp. 1011-1018 ◽

Cited By ~ 23

Author(s):

Michael H. Schiewe ◽

Jorge H. Crosa

Keyword(s):

Vibrio Anguillarum ◽

Virulence Plasmid ◽

Multicopy Plasmid ◽

Nick Translation ◽

Fish Pathogens ◽

Mole Percent ◽

Discrete Nature ◽

Sequence Relationships ◽

Restriction Endonuclease Cleavage

Examination of polynucleotide sequence relationships among 11 strains of Vibrio anguillarum biotype 2 isolated from moribund fish in North America and Japan demonstrated the highly conserved character of this group of fish pathogens and, moreover, confirmed its discrete nature from V. anguillarum biotype 1. Additional molecular analyses of the V. anguillarum biotype 2 strains revealed the universal presence of a multicopy plasmid with a molecular mass of approximately 20 × 106 daltons and a mole percent guanine plus cytosine of 44. The plasmid of strain DF3K was representative of this molecular species and was designated pMJ101. Subsequent DNA–DNA hybridizations using nick translation-labeled pMJ101 as a probe indicated all the 20 × 106 dalton plasmids were either identical or highly conserved, and, furthermore, that pMJ101 was apparently unrelated to either the virulence plasmid, pJM1, of V. anguillarum biotype 1 or to representatives of common plasmid incompatibility groups. The lack of relatedness between pMJ101 and pJM1 was further supported by differences in their restriction endonuclease cleavage patterns.

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