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Related Concept Videos

DNA Isolation01:24

DNA Isolation

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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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Related Experiment Video

Updated: Mar 21, 2026

Methylated DNA Immunoprecipitation
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Methylated DNA Immunoprecipitation

Published on: January 2, 2009

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UniClo technology exploits methylation for universal scarless DNA assembly.

Carol N Flores-Fernández1, Da Lin1, Katherine Robins1

  • 1Centre for Human Genetics, Nuffield Department of Medicine, University of Oxford, Oxford, OX3 7BN, United Kingdom.

Biology Methods & Protocols
|March 20, 2026
PubMed
Summary
This summary is machine-generated.

UniClo is a novel DNA assembly method that overcomes limitations of previous techniques. It enables scarless DNA assembly, even with internal restriction sites, offering greater flexibility for any DNA molecule.

Keywords:
DNA assemblydCas9/sgRNAmethylation-protectionrecombinant DNA methylasesscarless assembly vectorstype IIS restriction enzymes

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Biotechnology

Background:

  • Existing Golden Gate DNA assembly methods have limitations.
  • These include the need for sequence domestication, persistent scars, and multiple vectors/overhangs.

Purpose of the Study:

  • To develop a versatile and scarless DNA assembly technique.
  • UniClo aims to overcome the limitations of prior Golden Gate assembly methods.

Main Methods:

  • UniClo utilizes recombinant methylases to inactivate restriction enzyme sites.
  • CRISPR-dCas9 protects essential sites from methylation.
  • Engineered vectors trim overhangs to prevent scars.

Main Results:

  • UniClo allows assembly of DNA sequences with internal type IIS restriction sites.
  • Achieves fully scarless hierarchical assembly.
  • Requires only three assembly vectors and two universal overhang sequences.

Conclusions:

  • UniClo provides a flexible and efficient DNA assembly solution.
  • Enables the assembly of any DNA molecule, irrespective of sequence or application.
  • Overcomes key limitations of previous DNA assembly techniques.