Whole Genomic Amplification for Detecting Single-Gene Variants Causing Inherited Disease

The whole genome amplification (WGA) allows new clinical applications with minimal genetic material, such as in the genetic diagnosis of Mendelian diseases in embryos before implantation (Preimplantation Genetic Test for Mendelian Abnormalities, PGT-M). This approach allows couples to avoid the transmission of Mendelian disease by undergoing assisted reproduction treatment through in vitro fertilization (IVF). First, Preimplantation Genetic Testing for Aneuploidy (PGT-A) is used to identify chromosomal aneuploidies in IVF-generated embryos. Then, or in parallel, euploid embryos can be screened for specific diseases caused by variants in a single gene to achieve the conception of offspring free of a specific monogenic disease.Here, we detail the WGA preparation and two downstream usages: (1) preparation of PCR fragments for Sanger sequencing, exemplifying the diseases we detected for healthy embryo selection and transfer in IVF, and (2) detection of chromosome Y for embryo sex diagnosis.