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. 2026:2969:43-59.
doi: 10.1007/978-1-0716-4767-7_4.

Multiplexed Digital PCR for Evaluation of Karyotype Quality of Mouse Embryonic Stem Cells

Christy J Greenwood  1 , Alasdair J Allan  2 , Lydia Teboul  3
Affiliations

Multiplexed Digital PCR for Evaluation of Karyotype Quality of Mouse Embryonic Stem Cells

Christy J Greenwood et al. Methods Mol Biol. 2026.

Abstract

Digital PCR enables the high precision, absolute quantification of nucleic acid target sequences through volumetrically defined partitioning. This precision, coupled with the removal of PCR efficiency biases and the need for a standard curve, has instituted its widespread adoption over other quantitative methods. Digital PCR is particularly well suited for precise copy counting of chromosome numbers in cells, making it a viable alternative to traditional karyotyping in many instances. Here we describe a multicolor multiplex digital PCR method of screening ES cell clones for aneuploidy for cheaper and swifter validation. This method can also be easily applied to other in vitro cellular models that require accurate analysis for copy number variations.

Keywords: Aneuploidy; Digital PCR; Embryonic stem cells; Gene targeting; Karyotype; Multiplex assay.

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References

    1. Capecchi MR (1989) The new mouse genetics: altering the genome by gene targeting. Trends Genet 5:70–76
    1. Birling M-C, Yoshiki A, Adams DJ, Ayabe S, Beaudet AL, Bottomley J et al (2021) A resource of targeted mutant mouse lines for 5,061 genes. Nat Genet 53:416–419
    1. Nagy A, Gocza E, Diaz EM, Prideaux VR, Ivanyi E, Markkula M et al (1990) Embryonic stem cells alone are able to support fetal development in the mouse. Development 110:815–821. https://doi.org/10.1242/dev.110.3.815 - DOI - PubMed
    1. Liu X, Wu H, Loring J, Hormuzdi S, Disteche CM, Bornstein P et al (1997) Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission. Dev Dyn 209:85–91
    1. Longo L, Bygrave A, Grosveld FG, Pandolfi PP (1997) The chromosome make-up of mouse embryonic stem cells is predictive of somatic and germ cell chimaerism. Transgenic Res 6:321–328. https://doi.org/10.1023/A:1018418914106 - DOI - PubMed

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