Telomere length measurement-caveats and a critical assessment of the available technologies and tools.
Telomere length measurement-caveats and a critical assessment of the available technologies and tools.
复制标题
DOI：
10.1016/j.mrfmmm.2011.04.003
复制
发表时间：
2012-02-01
影响因子：
2.3
通讯作者：
Lansdorp, Peter M.
中科院分区：
文献类型：
--
作者：
Aubert, Geraldine;Hills, Mark;Lansdorp, Peter M.
端粒和端粒生物学的研究往往严重依赖于端粒DNA的检测和端粒重复序列长度的测量，无论是在单个细胞或细胞群体。有几种方法可以提供这类信息，通常不清楚哪种方法最适合解决特定的研究问题。需要考虑的主要变量是现有或可以提供的材料以及所需测量的准确性。本综述的目的是对最常用的方法进行全面总结，并讨论每种方法的优点和缺点。从基因组DNA开始的方法包括端粒限制性片段（TRF）长度分析、通过Q-PCR或MMQPCR相对于单拷贝基因的端粒重复序列的PCR扩增以及单端粒长度分析（STELA），单端粒长度分析是一种基于PCR的方法，其精确地测量来自个体染色体的端粒长度的全谱。另一套不同的方法依赖于荧光原位杂交（FISH）来检测单个细胞或染色体中的端粒重复序列。通过包括必要的校准步骤和适当的控制，这些方法可用于测量染色体和细胞中的端粒重复序列长度或含量。这些方法包括分别基于数字显微镜和流式细胞术的定量FISH（Q-FISH）和流式FISH。在这里，各种端粒长度测量方法的基本原理进行了描述，并强调其优点和缺点。本文还讨论了端粒长度分析的一些最新进展。在这篇评论中的信息应有助于选择最合适的方法来解决特定的研究问题，无论是在模式生物或人类受试者的端粒。
Studies of telomeres and telomere biology often critically rely on the detection of telomeric DNA and measurements of the length of telomere repeats in either single cells or populations of cells. Several methods are available that provide this type of information and it is often not clear what method is most appropriate to address a specific research question. The major variables that need to be considered are the material that is or can be made available and the accuracy of measurements that is required. The goal of this review is to provide a comprehensive summary of the most commonly used methods and discuss the advantages and disadvantages of each. Methods that start with genomic DNA include telomere restriction fragment (TRF) length analysis, PCR amplification of telomere repeats relative to a single copy gene by Q-PCR or MMQPCR and single telomere length analysis (STELA), a PCR-based approach that accurately measures the full spectrum of telomere lengths from individual chromosomes. A different set of methods relies on fluorescent in situ hybridization (FISH) to detect telomere repeats in individual cells or chromosomes. By including essential calibration steps and appropriate controls these methods can be used to measure telomere repeat length or content in chromosomes and cells. Such methods include quantitative FISH (Q-FISH) and flow FISH which are based on digital microscopy and flow cytometry respectively. Here the basic principles of various telomere length measurement methods are described and their strengths and weaknesses are highlighted. Some recent developments in telomere length analysis are also discussed. The information in this review should facilitate the selection of the most suitable method to address specific research question about telomeres in either model organisms or human subjects.
登录
查看更多内容
影响因子：
14.9
作者：
Cawthon RM
通讯作者：
Cawthon RM
影响因子：
11.4
作者：
BAIRD, DM;JEFFREYS, AJ;ROYLE, NJ
通讯作者：
ROYLE, NJ
影响因子：
30.8
作者：
Baird, DM;Rowson, J;Kipling, D
通讯作者：
Kipling, D
DOI：
10.1056/nejmra0903373
发表时间：
2009-12-10
期刊：
The New England journal of medicine
影响因子：
--
作者：
Calado RT;Young NS
通讯作者：
Young NS
影响因子：
2.7
作者：
Bryant, JE;Hutchings, KG;Griffith, JK
通讯作者：
Griffith, JK