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Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
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The hydration of cement takes place within the water-filled capillary pores. However, environmental elements can disrupt this process by evaporating water from the concrete surfaces. Sealed concrete with a water-cement ratio below 0.5 experiences self-desiccation, leading to water loss. The water loss in concrete is mitigated by curing. This technique involves keeping the concrete saturated to maintain the necessary temperature and moisture conditions, to optimally fill the spaces in the cement...
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Concrete members with a small surface-to-volume ratio are cured by oiling and moistening the forms before casting the concrete member. These forms can be left in place for a prolonged period to prevent moisture loss, and can be wetted if made of a material suitable for wetting. If the forms are removed early, the concrete member is moistened and covered with polythene sheets to maintain moisture. For large horizontal concrete surfaces exposed to dry weather, a temporary covering is suspended...
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Accelerating concrete curing is achieved by applying heat and additional moisture. This process accelerates the hydration of the cement, resulting in an earlier strength gain in the concrete. Steam curing is a method wherein the concrete products are either transported through a chamber on a conveyor belt or encased in plastic, allowing steam at atmospheric pressure to circulate freely around them. This process begins with a phase of moist curing that typically lasts between 3 to 5 hours, after...
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Related Experiment Video

Updated: Feb 10, 2026

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit
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Standardized Cloning and Curing of Plasmids.

Ida Lauritsen1, Se Hyeuk Kim1, Andreas Porse1

  • 1Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Lyngby, Denmark.

Methods in Molecular Biology (Clifton, N.J.)
|May 14, 2018
PubMed
Summary

This study presents standardized protocols for manipulating Standard European Vectors Architecture (SEVA) plasmids and a universal CRISPR-Cas9 system for plasmid removal, aiding synthetic biology research and bioengineering.

Keywords:
Cell factory designDNA assemblyPlasmidPlasmid curingSEVAVector backbone exchangepFREE

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Plasmids are essential for studying cells and gene expression in synthetic biology.
  • Standardized DNA vector handling is crucial for reproducible synthetic biology research.

Purpose of the Study:

  • To describe protocols for molecular cloning and genetic part exchange within the SEVA vector system.
  • To introduce a one-step CRISPR-Cas9 plasmid curing system (pFREE) for rapid bioengineering.
  • To demonstrate the application of pFREE for curing SEVA constructs.

Main Methods:

  • Development and validation of molecular cloning protocols for SEVA vectors.
  • Design and implementation of a universal CRISPR-Cas9-based plasmid curing system (pFREE).
  • Application of pFREE to remove SEVA constructs from engineered cells.

Main Results:

  • Established standardized protocols for SEVA vector manipulation, facilitating genetic part exchange.
  • Developed and validated the pFREE system for efficient, one-step plasmid curing.
  • Successfully demonstrated the curing of SEVA constructs using the pFREE system.

Conclusions:

  • The described SEVA protocols and the pFREE system provide valuable tools for synthetic biology.
  • These advancements enable faster testing and iterative bioengineering with diverse vector designs.
  • All developed vectors are available through SEVA/Addgene for broader research accessibility.