Relationship between fetal fraction and cfDNA concentration. A. cfDNA concentration was negatively correlated with fetal fraction in the sequencing of shorter cfDNA for NIPT. B. Among pregnant women in the same cfDNA concentration range, fetal fraction decreased with longer cfDNA fragment size and higher cfDNA concentration. C. The mean fetal fractions of maternal cfDNA concentrations < 0.121, 0.121-0.151, 0.152-0.181, 0.182-0.221, and > 0.221 ng/μl were 34.2%, 32.7%, 30.4%, 27.9%, and 27.1%, respectively. D. The mean fetal fractions of maternal plasma cfDNA concentrations < 0.121, 0.121-0.151, 0.152-0.181, 0.182-0.221, and > 0.221 ng/μl were 12.7%, 11.1%, 10.8%, 10.0%, and 8.7%, respectively. There was also a modest trend toward a lower fetal fraction with increasing cfDNA concentration in traditional NIPT (**P < 0.01).