用于检测小鼠胚胎G6pd基因特定CpG位点甲基化的聚合酶链反应

Polymerase chain reaction for the detection of methylation of a specific CpG site in the G6pd gene of mouse embryos.

作者信息

Zuccotti M, Grant M, Monk M

机构信息

Dipartimento Biologia Animale, University of Pavia, Italy.

出版信息

Methods Enzymol. 1993;225:557-67. doi: 10.1016/0076-6879(93)25036-2.

DOI:10.1016/0076-6879(93)25036-2

PMID:7694043

Abstract

This chapter describes the polymerase chain reaction (PCR) detection of methylation changes at specific CpG sites in DNA isolated from minute quantities of biological material, such as single preimplantation mouse embryos or small numbers of stem cells, or germ cells. We have concentrated on refining these techniques to monitor specific sites in X-linked genes for methylation changes associated with X-chromosome inactivation. The general principles of the HpaII-sensitive PCR assay described here should be applicable and adaptable to specific CpG sites in other genes of interest.

摘要

本章描述了聚合酶链反应（PCR）检测从微量生物材料（如单个植入前小鼠胚胎、少量干细胞或生殖细胞）中分离的DNA特定CpG位点甲基化变化的方法。我们专注于改进这些技术，以监测X连锁基因中的特定位点，检测与X染色体失活相关的甲基化变化。这里描述的HpaII敏感PCR检测的一般原理应适用于并可应用于其他感兴趣基因中的特定CpG位点。