Document Type

Honors Project

First Advisor

Dr. Moshe Khurgel

Degree Award Date

Spring 5-1-2021

Keywords

Axolotls, Sex-Typing, PCR, Real Time PCR, Real-Time PCR, qPCR

Disciplines

Cellular and Molecular Physiology | Molecular Genetics

Abstract

Sex-typing axolotls is useful in determining the role of sex on their physiology, gene expression and genome content. The traditional method of sex-typing through visual observations is not always reliable in this species. A more efficient and reliable method to determining the sex of an axolotl has been devised recently, which utilizes end-point polymerase chain reaction (PCR) based assay. In this study, we attempted to design a modified approach to sex-typing by using real-time PCR (qPCR). DNA was extracted from a limb or tail clipping and then amplified using qPCR. The gene of interest is located on the female sex chromosome, W, since female axolotls are ZW heterozygotes, while males are ZZ homozygotes. The control gene, E16E2, is specific to axolotls and was used to ensure the purity of sample DNA. The presence or absence of the sex gene was quantified via the detection of fluorescent probes on the amplified segments of DNA. After multiple trials, the control gene was detected in some of the samples and the presence or absence of the sex gene was noted. These data will be used to determine whether axolotls exhibit sexual dimorphism with respect to their rate of growth.

Recommended Citation

Leo, Cailyn. "Sex Determination in Axolotls Using PCR." Senior Honors Projects, Bridgewater College, 2021

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