一种基于液相探针捕获的Y染色体测序方法

• Format: Patent
• Language: Chinese
• Published: 22.03.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

一种基于液相探针捕获的Y染色体测序方法，包括如下步骤：S1.提取基因组DNA并片段化；S2.对片段化的DNA进行末端修复；S3.在DNA片段两端加上测序接头并纯化连接产物；S4.PCR扩增连有接头的片段DNA进行文库扩增并纯化；S5.文库质量控制；S6.探针区域的挑选及优化，并设计局部密集式探针；S7.将DNA文库和探针进行文库杂交反应，捕获目标DNA片段；S8.将捕获的目标DNA片段进行分离；S9.对分离后的目标DNA片段进行高通量测序。本发明利用液相探针捕获技术，通过对探针区域进行挑选与优化，对Y染色体的测序长度更长，就能够测试到更多的突变，实现了人类Y染色体20M区域的捕获测序。 The invention provides a Y chromosome sequencing method based on liquid phase probe capture. The method comprises the following steps of: S1, extracting genome DNA and fragmenting; S2, performing terminal repair on the fragmented DNA; S3, adding sequencing joints at two ends of the DNA fragment and purifying the connection product; S4, performing PCR amplification on the fragment DNA connected with the joints, and performing library amplification and purification; S5, controlling the library quality; S6, selecting and optimizing the probe area, and designing a local dense probe; S7, performinglibrary hybridization reaction on the DNA library and the probe, and capturing the target DNA fragment; S8, separating the capt