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Related Experiment Video

Updated: May 14, 2026

Plasmid-derived DNA Strand Displacement Gates for Implementing Chemical Reaction Networks
07:50

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Published on: November 25, 2015

RNA-friendly plasmid preparation.

Timothy W Nilsen

    Cold Spring Harbor Protocols
    |February 5, 2013
    PubMed
    Summary
    This summary is machine-generated.

    Plasmid DNA purification using miniprep kits can lead to RNase contamination, impacting experiments. Traditional cesium chloride (CsCl) density gradient methods offer a superior, RNase-free alternative for highly pure plasmid DNA.

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    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Genetics

    Background:

    • Miniprep and maxiprep kits are widely used for plasmid DNA preparation.
    • Plasmid DNA prepared with these kits is often used for in vitro transcription and cell transfection.
    • Contamination with RNases, particularly pancreatic RNase, is a significant issue with commercial DNA preparation kits.

    Purpose of the Study:

    • To highlight the problem of RNase contamination in plasmid DNA prepared using commercial kits.
    • To recommend an alternative purification method for obtaining highly pure plasmid DNA.
    • To provide a reliable protocol for RNase-free plasmid DNA suitable for sensitive downstream applications.

    Main Methods:

    • Evaluation of plasmid DNA purity from miniprep/maxiprep kits.

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  • Comparison with plasmid DNA purified using traditional cesium chloride (CsCl) density gradient ultracentrifugation.
  • Assessment of RNase contamination in DNA preparations.

    • Main Results:

    • Miniprep and maxiprep DNA frequently contains difficult-to-inactivate RNase contaminants.
    • Cesium chloride (CsCl) density gradient methods yield highly pure, RNase-free plasmid DNA.
    • The described CsCl protocol provides sufficient DNA for multiple experiments.

    Conclusions:

    • Traditional CsCl density gradient purification is recommended over commercial kits for plasmid DNA intended for transcription or transfection.
    • Investigative efforts should focus on RNase-free DNA purification to ensure experimental reproducibility.
    • The CsCl method provides a robust and scalable approach for obtaining high-quality plasmid DNA.