一种单染色体水平的甲基化测序方法

本发明涉及一种单染色体水平的甲基化测序方法，其包括以下步骤：(1)培养与收集待测样本中的淋巴细胞；(2)显微注射挑取单细胞并分离染色体以获得单倍体；(3)将单倍体进行稀释，用Bisulfite处理，将基因组中未发生甲基化的C碱基转换成U，进行PCR扩增后变成T，与原本具有甲基化修饰的C碱基区分，高通量测序技术进行甲基化测序。本发明提供的方法具有成本低、试验方法简单、耗时周期短以及设备成熟等优点，可广泛应用于癌症的早期诊断和鉴别诊断的临床研究中，其具备全基因组和单细胞水平甲基化检测方法的优势并可确定甲基化信息的遗传来源，从而可广泛应用于表观遗传的研究。

The invention relates to a methylation sequencing method of single-chromosome levels. The methylation sequencing method includes the steps of 1), culturing and collecting lymphocyte in to-be-detected samples; 2), selecting single cells with micro-injection and isolating chromosome to obtain monoploids; 3), releasing the monoploids and processing the same with Bisulfite, converting C base not subjected to methylation in genome into U which is changed into T after performing PCR (polymerase chain reaction) amplification, distinguishing from the C base originally having methylation modification, and utilizing the high-throughput sequencing technology to perform methylation sequencing detection. The methylation sequencing method has the advantages of low cost, simpleness in testing method, short time-consuming cycle, mature equipment and the like and can be widely used in clinical research of early diagnosis and differential diagnosis of cancers; the method has the advantages of methylation sequencing detection method of whole-genome and single cell levels, genetic origin of methylation information can be determined, and the method can be widely applied in research of epigenetic inheritance.