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Attachment of Sister Chromatids

Attachment of Sister Chromatids

As cells progress into mitosis, the nuclear envelope breaks down, and the condensed chromosomes are exposed to the array of bipolar microtubules of the mitotic spindle. The kinetochore, a large, disc-shaped protein complex, is present at the centromere region of the sister chromatids and acts as a binding site for the microtubules. Usually, the plus-end of a single microtubule is embedded within the kinetochore. However, some kinetochores first establish lateral contact with the side-wall of a...

Attachment of Sister Chromatids

Attachment of Sister Chromatids

As cells progress into mitosis, the nuclear envelope breaks down, and the condensed chromosomes are exposed to the array of bipolar microtubules of the mitotic spindle. The kinetochore, a large, disc-shaped protein complex, is present at the centromere region of the sister chromatids and acts as a binding site for the microtubules. Usually, the plus-end of a single microtubule is embedded within the kinetochore. However, some kinetochores first establish lateral contact with the side-wall of a...

Separation of Sister Chromatids

Separation of Sister Chromatids

At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...

Separation of Sister Chromatids

Separation of Sister Chromatids

At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...

Cohesins

Cohesins

Cohesin protein complexes are a molecular glue that holds two sister chromatids together. They play an important role both in mitosis and meiosis. In mitosis, all cohesin complexes present on the chromosomes are removed before the start of the anaphase stage.
Cohesin complexes in Meiotic Division
Meiosis involves two distinct rounds of chromosomal segregation and cell divisions— Meiosis I followed by Meiosis II – producing four daughter cells. Meiosis I includes the separation of homologous...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

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Related Experiment Video

Updated: Jul 5, 2026

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Published on: September 23, 2011

Analysis of sister-chromatid exchanges.

J German¹, B Alhadeff

¹New York Blood Center, New York, New York, USA.

Current Protocols in Human Genetics

|April 23, 2008

Summary

This study details a method for analyzing sister-chromatid exchanges (SCEs) in somatic cells. It uses bromodeoxyuridine (BrdU) incorporation and differential staining to visualize SCEs in proliferating lymphocytes.

Area of Science:

Cytogenetics
Molecular Biology
Cell Biology

Background:

Sister-chromatid exchanges (SCEs) are crucial indicators of DNA damage and repair.
Accurate cytogenetic analysis of SCEs requires actively proliferating cells and differentially stained sister chromatids.

Purpose of the Study:

To present a reliable protocol for the cytogenetic analysis of sister-chromatid exchanges (SCEs) in somatic cells.
To establish a method for differential sister chromatid staining using bromodeoxyuridine (BrdU) and specific staining techniques.

Main Methods:

Utilizing phytohemagglutinin (PHA)-stimulated blood lymphocyte cultures for cell proliferation.
Incorporating the thymidine analog bromodeoxyuridine (BrdU) into cellular DNA during replication.
Applying Hoechst 33258 staining, light exposure, heat treatment, and Giemsa staining to achieve differential chromatid visualization.

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Analysis of Nonhomologous End Joining and Homologous Recombination Efficiency in HEK-293T Cells Using GFP-Based Reporter Systems

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Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

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Related Experiment Videos

Last Updated: Jul 5, 2026

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

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Analysis of Nonhomologous End Joining and Homologous Recombination Efficiency in HEK-293T Cells Using GFP-Based Reporter Systems

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Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Published on: February 13, 2019

Main Results:

Successfully generated differentially stained metaphase chromosomes from BrdU-substituted cells.
Demonstrated the ability to recognize SCEs as reciprocal exchanges between sister chromatids.
Observed that bifilar BrdU-substituted chromatids exhibit a lighter purple stain compared to unifilarly substituted chromatids.

Conclusions:

The described protocol effectively enables the cytogenetic analysis of sister-chromatid exchanges (SCEs).
This method provides a robust approach for visualizing and quantifying SCEs in human somatic cells.
The differential staining technique is essential for accurate SCE detection and analysis in cytogenetic studies.