Whole-Genome Amplification on Single Circulating Trophoblast Cell

Whole-genome amplification (WGA) is a technique designed to amplify the entire genome from trace amounts of original template DNA, such as that from a single cell. This amplification provides sufficient DNA for downstream comprehensive genetic analysis. In cell-based noninvasive prenatal testing (cb-NIPT), single circulating trophoblast cells are isolated from the maternal bloodstream. Among the required steps in single trophoblast isolation are light paraformaldehyde fixation, permeabilization, and immunostaining, which are not compatible with all WGA methods. Following single trophoblast isolation and WGA, the amplified DNA is subjected to detailed SNP genotyping and next-generation sequencing for genetic testing. This protocol is a step-by-step guideline for WGA on circulating single trophoblasts isolated from maternal blood during pregnancy.