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Related Experiment Videos

Restriction generated oligonucleotides utilizing the two base recognition endonuclease CviJI*

N Swaminathan1, D George, K McMaster

  • 1CHIMERx, Madison, WI 53704.

Nucleic Acids Research
|April 25, 1994
PubMed
Summary

This study shows that the CviJI enzyme can convert anonymous DNA into specific oligonucleotide sequences. This enzymatic process has broad applications in molecular biology techniques.

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Area of Science:

  • Molecular Biology
  • Enzymology
  • Genetics

Background:

  • Restriction endonucleases are key tools in molecular biology.
  • CviJI is an unusual restriction endonuclease with flexible recognition sites.
  • DNA fragmentation is a critical step in many molecular biology workflows.

Purpose of the Study:

  • To investigate the enzymatic conversion of anonymous DNA into oligonucleotides using CviJI.
  • To characterize the DNA cleavage patterns of CviJI under relaxed conditions (CviJI*).
  • To explore the potential applications of CviJI-generated fragments in molecular biology.

Main Methods:

  • Enzymatic digestion of DNA using CviJI and CviJI* under specific reaction conditions.
  • Analysis of DNA fragment sizes produced by CviJI* restriction of pUC19.

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  • Thermal denaturation of DNA fragments to generate sequence-specific oligonucleotides.

    • Main Results:

    • CviJI* produced DNA fragments predominantly in the 18-56 bp range, deviating from theoretical predictions.
    • 96% of CviJI* fragments were within the 18-56 bp size range.
    • Thermal denaturation yielded sequence-specific oligonucleotides homologous to the DNA template.

    Conclusions:

    • CviJI enzyme facilitates the enzymatic conversion of anonymous DNA into sequence-specific oligonucleotides.
    • The characterized CviJI* activity offers novel possibilities for DNA manipulation.
    • This enzymatic approach has significant implications for both established and new molecular biology procedures.