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Related Experiment Video

Updated: Jul 10, 2025

Kinetic Measurement and Real Time Visualization of Somatic Reprogramming

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Repli-seq Sample Preparation using Cell Sorting with Cell-Permeant Dyes.

Silvia Meyer-Nava^1,2, Anala V Shetty^1,2, Juan Carlos Rivera-Mulia^1,2,3

¹Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota Medical School, Minneapolis, Minnesota.

Current Protocols

|November 27, 2023

Summary

This summary is machine-generated.

This study presents optimized Repli-seq methods for live cells and intact nuclei. These new protocols reduce cell input, processing time, and sample loss while maintaining high-quality replication timing data.

Keywords:

Repli-seq cell cycle flow cytometry intact nuclei replication timing

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Published on: October 26, 2018

Area of Science:

Molecular Biology
Genomics
Epigenetics

Background:

Replication timing correlates with gene expression and chromatin organization.
It changes during cell differentiation and is altered in disease.
Genome-wide replication timing analysis uses Repli-seq, typically requiring large numbers of fixed cells.

Purpose of the Study:

To develop optimized Repli-seq methods for live cells and intact nuclei.
To overcome limitations of current fixation-dependent protocols.
To enable analysis of replication timing in sensitive or low-cell-number samples.

Main Methods:

Optimized protocols for processing live cells and intact nuclei for Repli-seq.
Methods for isolation, staining, and processing of nuclei from difficult-to-dissociate samples.
Comparison of Repli-seq data from live cells/nuclei with standard protocols.

Main Results:

Successful implementation of Repli-seq using live cells and intact nuclei.
Reduced cell input, processing time, and sample loss compared to standard methods.
High-quality Repli-seq data comparable to conventional protocols.

Conclusions:

Optimized live-cell and intact-nuclei Repli-seq protocols are effective.
These methods expand the applicability of Repli-seq to challenging sample types.
The new protocols provide high-quality replication timing data with improved efficiency.