Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Types of Selection01:46

Types of Selection

40.2K
Natural selection influences the frequencies of particular alleles and phenotypes within populations in several different ways. Primarily, natural selection can be directional, stabilizing, or disruptive. Directional selection favors one extreme trait and shifts the population towards that phenotype while selecting against individuals displaying alternate traits. Stabilizing selection favors an intermediate trait with a narrow range of variation. Deviation from the optimal phenotype towards an...
40.2K
Catalytically Perfect Enzymes01:07

Catalytically Perfect Enzymes

3.9K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
 
Most enzymes...
3.9K
Gene Evolution - Fast or Slow?02:05

Gene Evolution - Fast or Slow?

7.0K
The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...
7.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

LiraSearch-ultrafast ligand shape and electrostatic matching server.

Bioinformatics advances·2026
Same author

Quantifying and Minimizing the Variance of Gradient Insulator-Based Dielectrophoresis.

Micromachines·2026
Same author

Are we there yet with XNA aptamers?

RSC advances·2026
Same author

A single-cell cytokine dictionary of human peripheral blood.

Research square·2026
Same author

A single-cell cytokine dictionary of human peripheral blood.

bioRxiv : the preprint server for biology·2025
Same author

Deciphering coulombic loss in lithium-ion batteries and beyond.

Nature communications·2025
Same journal

Editorial: Epigenetic and genetic mechanisms underlying cardiovascular diseases and neurodevelopmental disorders, volume II.

Frontiers in molecular biosciences·2026
Same journal

Integrated transcriptomic profiling reveals oncogenic pathways and chimeric transcripts in equine sarcoid lesions with predominant BPV1 detection.

Frontiers in molecular biosciences·2026
Same journal

Mesenchymal stem cells-derived extracellular vesicles as a novel drug delivery carrier: engineering strategies and clinical safety estimation.

Frontiers in molecular biosciences·2026
Same journal

Preparation and analysis of tobacco glycosides, and the relationship between glycoside aglycones and pyrolysis products: a review.

Frontiers in molecular biosciences·2026
Same journal

Peritoneal metastasis in pancreatic cancer: molecular mechanisms, microenvironmental remodeling, and emerging intraperitoneal interventions.

Frontiers in molecular biosciences·2026
Same journal

Insights from LC-MS-based cerebrospinal fluid metabolomics in tuberculous meningitis.

Frontiers in molecular biosciences·2026
See all related articles

Related Experiment Video

Updated: Jun 9, 2025

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

30.5K

Navigating directed evolution efficiently: optimizing selection conditions and selection output analysis.

Paola Handal-Marquez1, Hoai Nguyen1, Vitor B Pinheiro1

  • 1Department of Pharmaceutical and Pharmacological Sciences, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.

Frontiers in Molecular Biosciences
|October 23, 2024
PubMed
Summary
This summary is machine-generated.

Directed evolution optimizes polymerase engineering by analyzing selection conditions and sequencing coverage. This method enhances the efficiency of creating novel DNA and xenobiotic nucleic acid (XNA) polymerases for biotechnology.

Keywords:
design of experimentsdirected evolutionfitness landscapenext-generation sequencing (NGS) data analysispolymerase engineering

More Related Videos

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.2K
Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
10:50

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

10.9K

Related Experiment Videos

Last Updated: Jun 9, 2025

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

30.5K
In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.2K
Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
10:50

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

10.9K

Area of Science:

  • Biochemistry and Molecular Biology
  • Enzyme Engineering
  • Synthetic Biology

Background:

  • Directed evolution enables protein variant isolation despite incomplete sequence-function understanding.
  • Xenobiotic nucleic acid (XNA) polymerases are crucial for therapeutic and biotechnological applications but currently exhibit lower efficiency and fidelity than natural counterparts.
  • Existing directed evolution methods for polymerase engineering lack comprehensive analysis of selection and library biases.

Purpose of the Study:

  • To develop a method for analyzing the impact of selection conditions on the success and efficiency of directed evolution for DNA and XNA polymerase engineering.
  • To investigate the influence of selection conditions on polymerase fidelity at both population and individual mutant levels.
  • To determine optimal sequencing coverage requirements for identifying significantly enriched mutants in directed evolution experiments.

Main Methods:

  • Focused analysis on the directed evolution pipeline for DNA and XNA polymerase engineering.
  • Evaluation of selection conditions' effects on selection success, efficiency, and fidelity.
  • Exploration of sequencing coverage needs specific to directed evolution, differentiating from other -omics approaches.
  • Identification of sequencing coverage thresholds for accurate mutant identification.

Main Results:

  • A methodology was established to understand how selection conditions impact directed evolution outcomes.
  • The study identified critical sequencing coverage thresholds for precise mutant enrichment detection.
  • Optimized selection protocols using smaller libraries and cost-effective next-generation sequencing (NGS) were demonstrated.

Conclusions:

  • The developed methodology streamlines directed evolution processes for polymerase engineering.
  • Insights into selection parameters and sequencing coverage enhance the effectiveness of enzyme evolution strategies.
  • This approach is broadly applicable to directed evolution of enzymes beyond polymerases.