STR analysis and Prenatal Sex Genotyping by PCR of Fetal Cell-Free DNA purified from 18 ul of a 10-week pregnant woman's bloodPCR Success Story #24
Welcome to our lab mini16 miniPCR!
This PCR Success Story was entirely performed using the mini16 thermocycler from miniPCR & Amplyus
Confirmed : it's a BOY !
Non-specific AMEL Y amplification from women’s DNA was caused by the 3′-5′ exonuclease activity of high-fidelity DNA polymerases.
Because both X and Y-specific amelogenin PCR reactions use the same reverse primer, the HiFi DNA polymerases generate a lot of ssDNA strands using the Rxy primer. Hence, increasing amounts of ssDNA amplified from the women’s X chromosome favored the binding of the AMEL Y forward primer thanks to high levels of ‘fidelity’ inherent to 3′-5′ exo activity. See the first section for AMEL X and Y sequence mismatches.
Genuine Taq DNA Polymerases are devoid of 3′-5′ exonuclease activity. Hence, they are more suitable for genotyping in general because they amplify from primer-specific DNA sequences and mismatches at the 3′ end of DNA primers cannot be chewed back!