Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Direct end labelling of telomeres

A Kolchinsky1, P M Gresshoff

  • 1University of Tennessee, Knoxville 37901-1071.

Genome
|April 1, 1993
PubMed
Summary

Researchers developed a new method for direct telomere end labeling using T4 DNA polymerase. This technique successfully labeled telomeres in yeast and soybean, revealing length polymorphisms in soybean subspecies.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Development and differentiation of haploid Lycopersicon esculentum (tomato).

Planta·2014
Same author

Transfer of organelles of the alga Chlamydomonas reinhardii into carrot cells by protoplast fusion.

Planta·2014
Same author

Viability of Rhizobium bacteroids isolated from soybean nodule protoplasts.

Planta·2014
Same author

Cycloheximide resistance in carrot culture.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·2013
Same author

Ecotypic variation of in vitro plantlet formation in white clover (Trifolium repens).

Plant cell reports·2013
Same author

Rapid derepression of in vitro nitrogenase activity in a Rhizobium strain which nodulates legumes and the nonlegume Parasponia.

Plant cell reports·2013

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Telomeres, the protective caps of chromosomes, are crucial for genomic stability.
  • Accurate methods for telomere analysis are essential for understanding chromosome dynamics and genetic variation.
  • Previous telomere labeling techniques often involved indirect methods or lacked resolution.

Purpose of the Study:

  • To present a novel, direct end-labeling method for telomeres.
  • To validate the technique using yeast chromosomes and assess its applicability to plant genomes.
  • To determine telomere length and identify polymorphisms in soybean subspecies.

Main Methods:

  • Utilized T4 DNA polymerase to generate single-stranded 5' ends of telomeres in agarose-embedded, chromosome-sized DNA.
  • Employed direct enzymatic labeling with [alpha-32P]dGTP.
  • Separated labeled DNA using pulsed-field gel electrophoresis (PFGE) and two-dimensional gel electrophoresis for detailed analysis.

Main Results:

  • Successfully labeled telomeres in yeast chromosomes, with labeled DNA maintaining integrity after PFGE.
  • Generated a fingerprint-like pattern of labeled telomeres in yeast, comparable to hybridization results.
  • Applied the method to soybean (Glycine max and Glycine soja), enabling telomere size determination and revealing length polymorphisms between subspecies.

Conclusions:

  • The direct end-labeling method provides a robust approach for telomere analysis.
  • This technique is effective for studying telomere length and variation in diverse organisms, including plants.
  • The findings facilitate comparative genomics and the study of genetic diversity in soybean.

Related Experiment Videos