A method for gender test of mouse haploids by polymerase chain reaction

Abstract

Objective To establish a polymerase chain reaction (PCR) method with a high sensitivity for gender test of single mouse sperm and oocyte. Methods PCR system was set up to amplify repeat sequences on Rhox or Rbmy gene of the sex chromosome in a single sperm or an oocyte, and the gender was determined by using the differences of melting curves of amplification in PCR or nucleotide bands separated by agarose gel electrophoresis. Results The PCR system could reliably amplify the repeat sequences on Rhox or Rbmy gene in a single sperm or an oocyte, it can amplify haploid DNA in single sperm cell (PCR detection rate was 75%) and oocyte (PCR detection rate was 100%) when compared with the traditional method, which could be used for gender test. Conclusion By amplifying the repeat sequences on the sex chromosomes, the PCR method is capable to be used for gender test of a single sperm with the high sensitivity, which could be also applied for early embryos, such as blastula. Key words: Gender test; Mouse; Haploid; Multiple polymerase chain reaction