Detection of FecB gene polymorphisms in sheep using rapid and low-cost tetra-ARMS PCR

Document Type : Research Paper

Authors

1 Ph.D student of genetics & animal Biotechnology, Department of Animal science, Ferdowsi University of Mashhad Khorasan Razavi - Iran

2 Associate Professor of Animal Genetics and Breeding, Khorasan Razavi Agricultural and Natural Resources Research and Education center, AREEO, Mashhad, Iran

3 Ph.D student of genetics & animal biotechnology, Departman of animal science, Ferdowsi university of Mashhad,, Khorasan Razavi , Iran.

Abstract

The FecB gene is one one of the major genes associated with the twinning trait in sheep. AvaII enzyme has a restriction site containing FecB gene mutation is used for genotyping by PCR-RFLP method. Due to toxicity and expensive price, using this enzyme is not considered reasonable and economical. So, this research was performed to design specific primers for rapid and economical detection of FecB gene. Tetra ARMs PCR technique, caucuses elimination of AvaII enzyme, replacement of acrylamide gel by safe agarose gel, and of course saving time and money for genotyping. Four primers were designed, based of Tetra ARMs PCR method, and were tested on 3000 samples. After PCR and electrophoresis, three fragments with a length of 108, 213 and 276 bp were amplified and finally three genotypes were observed for FecB gene. In order to validate the designed primers, 20 samples were randomly sequenced. After validation, the result of this research indicates that the final obtained genotype in Tetra ARMs PCR method is same as the results from PCR-RFLP method. Tetra ARMs PCR method is rapid and economical for detection of FecB in a larger number of samples and can replace by enzyme digestion method which is all of more expensive.

Keywords

References

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Animal Sciences Journal
Volume 34, Issue 133
March 2022
Pages 147-156

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