Objective: To investigate whether the mitochondrial DNA content could predict the embryo viability Design: Retrospective analysis. Setting: Reproductive genetics laboratory Patient(s): A total of 421 biopsied samples obtained from 129 patients Intervention(s): Embryo biopsies samples underwent whole genome amplification (WGA) and were tested by next generation sequencing (NGS) and array Comparative Genomic Hybridization (aCGH), 30 samples were selected randomly to undergo quantitative real-time polymerase chain reaction (qPCR). Main Outcome Measure(s): Those embryos which obtained the consistent chromosome status determined both aCGH and NGS platform were further classified.We investigated the relationship of mtDNA content with several factors including female patient age, embryo morphology, chromosome status, and live birth rate of both blastocysts and blastomeres. Result(s):A total of 386 (110 blastomeres and 276 blastocysts) out of 399 embryos showed consistent chromosome status outcome. We found no statistically difference was observed in aneuploid and euploid blastocysts (p=0.14), the same phenomenon was observed in aneuploid and euploid blastomeres (p=0.89). Similarly, the mtDNA content was independent of female patient age, embryo morphology and live birth rate. Conclusion(s):The mtDNA content did not provide a reliable prediction of the viability of blastocysts to initiate a pregnancy.