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Restriction Enzymes01:11

Restriction Enzymes

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Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
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Base Excision Repair01:54

Base Excision Repair

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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...
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Related Experiment Video

Updated: Jul 26, 2025

Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor
09:33

Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor

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Using Exonucleases for Aptamer Characterization, Engineering, and Sensing.

Obtin Alkhamis1, Juan Canoura1, Phuong T Ly1

  • 1Department of Chemistry, North Carolina State University, 2620 Yarbrough Dr., Raleigh, North Carolina 27695, United States.

Accounts of Chemical Research
|June 14, 2023
PubMed
Summary
This summary is machine-generated.

Enzyme-assisted methods simplify aptamer engineering and characterization. Nucleases enhance aptamer sensors, enabling label-free detection and high-throughput screening for diverse applications.

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Kinetic Screening of Nuclease Activity using Nucleic Acid Probes
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Last Updated: Jul 26, 2025

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Kinetic Screening of Nuclease Activity using Nucleic Acid Probes
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Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Published on: November 1, 2019

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Aptamers are nucleic acid-based biorecognition elements with potential in diagnostics and monitoring.
  • Current challenges include limited sensitivity, complex characterization, and high costs in aptamer development.
  • Nuclease enzymes offer a promising avenue to overcome these limitations.

Purpose of the Study:

  • To develop novel enzymatic methodologies for aptamer engineering and characterization.
  • To enhance the sensitivity and streamline the development of aptamer-based sensors.
  • To create high-throughput methods for aptamer screening and affinity determination.

Main Methods:

  • Utilized exonuclease digestion inhibition upon aptamer-ligand binding.
  • Developed structure-switching aptamers via single-step exonuclease truncation.
  • Established a label-free detection platform using direct aptamer selection outputs.
  • Employed exonucleases for high-throughput aptamer affinity and specificity characterization.

Main Results:

  • Simplified aptamer engineering through single-step truncation.
  • Achieved nanomolar-level analyte detection with ultralow background in biological samples.
  • Enabled multiplexed detection using molecular beacons.
  • Facilitated comprehensive aptamer analysis and identification of enhanced aptamers.
  • Demonstrated high-throughput screening of aptamer candidates and aptamer-ligand pairs.

Conclusions:

  • Enzymatic technologies significantly streamline aptamer characterization and sensor development.
  • Novel methods address key challenges in aptamer engineering and application.
  • Potential for rapid identification of optimal aptamers for specific applications with automation.