Veriseq pgs workflow

The PGT-A protocol was similar to our previous report [33 (link)]. In brief, genomic DNA was extracted and amplified using the SurePlex DNA Amplification System (Illumina, San Diego, CA, USA). The amplified DNA product was used to prepare the genomic DNA libraries according to the VeriSeq PGS workflow (Illumina, USA). BlueFuse Multi Software (Illumina, USA) was used for data analysis, and the diploid–aneuploid levels of each sample were examined by at least two technicians. Based upon diploid–aneuploid mosaic ratios measured using the hr-NGS platform for biopsied cells [36 (link),37 (link),38 (link)], blastocysts were classified into the following three groups: (i) euploid blastocysts with mosaicism levels ≤ 20%; (ii) mosaic blastocysts with mosaicism levels between 20% and 50%; and (iii) aneuploid blastocysts with mosaicism levels > 50%.

The PGT-A protocol employed in this study was similar to that described in our previous report [29 (link)]. Genomic DNA was extracted and amplified using the SurePlex DNA Amplification System (Illumina, San Diego, CA, USA). The amplified DNA product was utilized to generate genomic DNA libraries following the VeriSeq PGS workflow (Illumina, USA). Data analysis was performed using the BlueFuse Multi Software (Illumina, USA), and at least two technicians assessed the diploid–aneuploid levels of each sample. Blastocysts were categorized into three groups based on diploid–aneuploid mosaic ratios determined using the high-resolution next generation sequencing (hr-NGS) platform on biopsied cells [31 (link),32 (link),33 (link)]: (i) euploid blastocysts with mosaicism levels ≤ 20%; (ii) mosaic blastocysts with mosaicism levels between >20% and <80%; and (iii) aneuploid blastocysts with mosaicism levels ≥ 80%.