A Real-Time Quantitative PCR Protocol for the Quantification of Plasmid Copy Number in Lactococcus lactis

Sofia O D Duarte; Gabriel A Monteiro

doi:10.1007/978-1-0716-3358-8_12

A Real-Time Quantitative PCR Protocol for the Quantification of Plasmid Copy Number in Lactococcus lactis

Methods Mol Biol. 2023:2967:151-157. doi: 10.1007/978-1-0716-3358-8_12.

Authors

Sofia O D Duarte^{1

2}, Gabriel A Monteiro^{3

4}

Affiliations

¹ iBB- Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
² Associate Laboratory i4HB-Institute for Health and Bioeconomy at Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
³ iBB- Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal. [email protected].
⁴ Associate Laboratory i4HB-Institute for Health and Bioeconomy at Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal. [email protected].

PMID: 37608109
DOI: 10.1007/978-1-0716-3358-8_12

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.

Keywords: Lactococcus lactis; Plasmid copy number; Real-time quantitative PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA Copy Number Variations
Lactococcus lactis* / genetics
Plasmids / genetics
Real-Time Polymerase Chain Reaction