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Updated: Jul 29, 2026
Accurate and Phenol Free DNA Sexing of Day 30 Porcine Embryos by PCR
Published on: February 14, 2016
An accurate and rapid gender determination assay in single cells by the capillary polymerase chain reaction method
T Hashiba1, K Sueoka, M Kuroshima
1Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan.
Purpose:
In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary polymerase chain reaction (PCR) method using rapid thermal cycling programs to determine the gender of single amniocytes.
Methods:
Single amniocytes from each amniotic fluid sample were isolated by micromanipulation and their gender was determined by a multiplex PCR assay in a capillary tube, using primers that amplify a 308-bp DXZ1 and a 154-bp DYZ1 repeat sequence on the X and Y chromosomes, respectively.
Results:
All four thermal cycling programs, which took 180, 150, 120, and 90 min, were 100% accurate in diagnosing the gender of single amniocytes. No DNA contamination was observed in any samples.
Conclusions:
The multiplex PCR assay was rapid and accurate in diagnosing gender in single cells and may be clinically applicable in PGD.
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