Authors:Guo Chen and Xingming Deng

作者：Guo Chen 和 Xingming Deng

Journal:Bio-Protocol, 2018, Volume 8, Issue 17

期刊：Bio-Protocol, 2018年, 第8卷, 第17期

DOI:10.21769/BioProtoc.2994

Abstract:

This paper details a protocol for synchronizing cell populations using the double thymidine block method. Thymidine, a nucleoside analog, is used to arrest cells at the G1/S boundary, thereby synchronizing the cell cycle. This technique is essential for studies focusing on cell cycle regulation, DNA replication, and mitosis.

摘要：

本文详细介绍了一种使用双胸苷阻滞方法实现细胞群体同步化的方案。胸苷是一种核苷类似物，用于将细胞阻滞在G1/S边界，从而实现细胞周期的同步化。这一技术对于研究细胞周期调控、DNA复制和有丝分裂至关重要。

Key Components and Steps:

1.Thymidine Treatment:

Thymidine is added to the cell culture medium to inhibit DNA synthesis.

The first thymidine block lasts for 18 hours, followed by a release period to allow cells to progress through the S phase.

A second thymidine block is then applied to ensure synchronization.

2.Cell Cycle Analysis:

After the second block, cells are released into fresh medium, and samples are collected at different time points.

Flow cytometry is used to analyze cell cycle progression.

3.Applications:

This method is used to study various aspects of cell cycle regulation, DNA damage response, and cellular processes tied to specific cell cycle phases.

4.Materials and Reagents:

Thymidine (CAS Number: 50-89-5) is the primary reagent used in this protocol.

Cell culture media and supplements necessary for maintaining and growing the cells.

主要步骤和成分：

1.胸苷处理:

将胸苷加入细胞培养基中以抑制DNA合成。

第一次胸苷阻滞持续18小时，然后释放一段时间以允许细胞通过S期。

随后进行第二次胸苷阻滞以确保同步化。

2.细胞周期分析:

在第二次阻滞后，将细胞释放到新鲜培养基中，并在不同时间点收集样本。

使用流式细胞术分析细胞周期进程。

3.应用:

该方法用于研究细胞周期调控、DNA损伤反应以及与特定细胞周期阶段相关的细胞过程。

4.材料和试剂:

胸苷（CAS号：50-89-5）是此协议中使用的主要试剂。

维持和培细胞所需的细胞培养基和补充剂。

Key Findings:

• The double thymidine block method effectively synchronizes cells at the G1/S boundary.
• It is a reliable and reproducible technique suitable for various cell types.
• This method allows researchers to obtain a population of cells at similar stages of the cell cycle, facilitating the study of cell cycle-specific processes.

主要发现：

• 双胸苷阻滞方法有效地将细胞同步化至G1/S边界。
• 这是一种适用于各种细胞类型的可靠且可重复的方法。
• 该方法使研究人员能够获得处于细胞周期相似阶段的细胞群体，方便研究细胞周期特定过程。

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