DNA methylation protocol for analyzing cell-free DNA in the spent culture medium of human preimplantation embryos

被引：5

作者：

Gao, Yuan ^{[1
,2
,3
]}

Chen, Yidong ^{[1
,2
,4
,5
]}

Qiao, Jie ^{[1
,2
,3
,4
,5
]}

Huang, Jin ^{[1
,2
,4
,5
]}

Wen, Lu ^{[1
,2
,4
]}

机构：

[1] Peking Univ, Hosp 3, Sch Life Sci, Biomed Pioneering Innovat Ctr,Dept Obstet & Gyneco, Beijing 100871, Peoples R China

[2] Peking Univ, Hosp 3, Beijing Adv Innovat Ctr Genom, Ctr Reprod Med, Beijing 100871, Peoples R China

[3] Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing 100871, Peoples R China

[4] Minist Educ, Key Lab Assisted Reprod, Key Lab Cell Proliferat & Differentiat, Beijing 100871, Peoples R China

[5] Beijing Key Lab Reprod Endocrinol & Assisted Repr, Beijing 100871, Peoples R China

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 02期

基金：

中国国家自然科学基金;

关键词：

Bioinformatics; Cell Biology; Sequence Analysis; Sequencing;

D O I：

10.1016/j.xpro.2023.102247

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Cell-free DNA (cfDNA) in spent embryo culture media (SECM) provides prospects for noninvasive preimplantation genetic testing. Here, we present a post-bisulfite-adapter-tagging (PBAT)-based whole-genome DNA methylation sequencing protocol (SECM-PBAT) for human SECM cfDNA analysis. We describe steps for SECM lysis, bisulfite conversion and purification, preamplification by random priming, tagging adapter II, and library establishment. We then detail library quality control, sequencing, and bioinformatics analysis. This approach simultaneously detects chromosome aneuploidy and deduces the proportional contributions of cellular components. For complete details on the use and execution of this protocol, please refer to Chen et al. (2021).1

引用

页数：21

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