Abstract
The determination of the number of plasmid copies in each cell of Lactococcus lactis is critical for the control and regulation of the production of recombinant proteins and plasmids. This protocol describes a method for the determination of the plasmid copy number per genome of L. lactis, which is based on the detection by real-time quantitative PCR of the number of plasmid molecules and the number of chromosomes and subsequently their ratio after calculating the amplification efficiency.
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References
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Acknowledgments
This work was supported by FCT-Portuguese Foundation for Science and Technology (project grant PTDC/BTM-SAL/28624/2017) and funding received from FCT, Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDB/04565/2020 and UIDP/04565/2020 of the Research Unit Institute for Bioengineering and Biosciences, iBB, and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB.
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Duarte, S.O.D., Monteiro, G.A. (2023). A Real-Time Quantitative PCR Protocol for the Quantification of Plasmid Copy Number in Lactococcus lactis. In: Domingues, L. (eds) PCR. Methods in Molecular Biology, vol 2967. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3358-8_12
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DOI: https://doi.org/10.1007/978-1-0716-3358-8_12
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3357-1
Online ISBN: 978-1-0716-3358-8
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