Multiplexing: An Efficient Way of Genetic Monitoring of Laboratory Mice using Microsatellite Markers

DOI: 10.18805/ag.D-5470    | Article Id: D-5470 | Page : 358-364
Citation :- Multiplexing: An Efficient Way of Genetic Monitoring of Laboratory Mice using Microsatellite Markers.Agricultural Science Digest.2022.(42):358-364
Azmat Naseem, Shashi Ahire, Arvind Ingle aingle@actrec.gov.in
Address : Laboratory Animal Facility, Cancer Research Institute-Advanced Centre for Treatment, Research and Education in Cancer, Navi Mumbai-410 210, Maharashtra, India.
Submitted Date : 4-08-2021
Accepted Date : 24-02-2022


Background: Inbred strains of laboratory mice have been widely used in biomedical research. An inbred strain is the one which has been maintained by sibling mating for twenty or more consecutive generations. Except for the sex difference, an inbred strain is homozygous at all loci. Most of the traits are consistent, do not vary in offspring and respond to experimental procedures uniformly. This enables researchers to produce reliable data for therapeutic purposes, hence inbred strain must be genetically pure over the generation.
Methods: A total of 20 mice samples from 10 inbred strains i.e., A/J, BALB/c, C3H/J, CD1, C57BL/6, DBA/2, FVB/NJ, ICRC, Swiss Webster and Swiss/Ba were screened for their genetic purity with the help microsatellite markers. Tail DNA was isolated using Proteinase/K-Phenol-chloroform extraction method and quantified by Nanodrop. DNA obtained was used for microsatellite marker analysis by standardizing multiplex-PCR. Seven multiplex (duplex) panels were successfully established by optimizing various reaction conditions to analyse any deviation in the genetic profile. 
Result: Upon comparing genetic profiles obtained in multiplex PCR with the information available on Mouse Genome Informatics database, it was concluded that most of the inbred mice are genetically consistent as they showed zero divergence in observed base pair size mainly due to stringent breeding protocol and optimum living conditions. The work done also signifies a cost-effective, time saving, high throughput and robust method of genetic monitoring of laboratory mice strains.


Genetic monitoring Mice Microsatellite markers Multiplex PCR


  1. Abdul-Muneer, P.M. (2014). Application of microsatellite markers in conservation genetics and fisheries management: recent advances in population structure analysis and conservation strategies. Genetic Research International. Article ID 691759: 11 pages. http://dx.doi.org/10.1155/ 2014/691759.
  2. Benavides, F. Rulicke, T. Prins, J.B. Bussell, J. Scavizzi, F. Cinelli, P. Herault, Y. Wedekind, D. (2020). Genetic quality assurance and genetic monitoring of laboratory mice and rats: FELASA Working Group Report. Laboratory Animals. 54(2): 135-48.
  3. Bryda, E.C. and Riley, L.K. (2008) Multiplex microsatellite marker panels for genetic monitoring of common rat strains. Journal of American Association of Laboratory Animal Science. 47(3): 37-41.
  4. Ghatak, S. Muthukumaran, R.B. Nachimuthu, S.K. (2013). A simple method of genomic DNA extraction from human samples for PCR-RFLP analysis. Journal of Biomolecular Techniques. 24(4): 224.
  5. Guenet, J.L. and Benavides, F.J. (2010) Genetic Monitoring of Laboratory Rodents. In: Molecular Diagnostics, [(Eds.) George P Patrinos and Wilhelm Ansorge], Academic Press, Oxford, 2nd edition, 461-469.
  6. Ingle, A.D. and Ahire, S.D. (2013). Checking genetic integrity of laboratory mice with the help of conventional methods. Research Animals. 1(1): 47- 55.
  7. Han, F. Wang, J. Ding, L. Hu, Y. Li, W. Yuan, Z. Guo, Q. Zhu, C. Yu, L. Wang, H. Zhao, Z., Jia, L. Li, J. Yu, Y. Zhang, W. Chu, G. Chen, S. Li, B. (2020). Tissue engineering and regenerative medicine: Achievements, future and sustainability in Asia. Frontiers in Bioengineering and Biotechnology. 8: 83. DOI=10.3389/fbioe.2020.00083.
  8. Litt, M. and Luty, J.A. (1989). A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. American Journal of Human Genetics. Mar. 44(3): 397.
  9. Perez-Jimenez, M. Besnard, G. Dorado, G. Hernandez, P. (2013). Varietal tracing of virgin olive oils based on plastid DNA variation profiling. PLoS One. 8(8): e70507.
  10. Thorat, R.A. and Ingle, A.D. (2012). An attempt of cryopreservation of mouse embryo at the ACTREC Laboratory Animal Facility in India. Experimental Animals. 61(2): 139-145.
  11. Thorat, R.A. and Ingle, A.D. (2017). Cryopreservation of mouse embryo using vitrification method. Journal of Laboratory Animal Science. 4(2): 7-12.
  12. Thorat, R.A. and Ingle, A.D. (2020). Breeding performance with special reference to pre-weaning mortality of C57BL/6 and B6D2F1 hybrid strain of mice maintained at ACTREC Animal Facility, Journal of Laboratory Animal Science. 6(2): 19-23.
  13. Thorat, R.A. and Ingle, A.D. (2021). Tackling the Covid-19 pandemic in Animal Facilities: ACTREC perspectives. Agricultural Review. 42(2): 203- 208.
  14. Thorat, R.A., Ahire, S.D. and Ingle, A.D. (2013). Re-establishment of breeding colony of NOD SCID mice from revival of cryo-preserved embryos. Lab Animals (Asia Pacific), 42(4): 131-134.

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