A Simple Technique for Isolating Plasmids from Escherichia Coli for Efficient Chemical Transfection of Human Cell Culture

Authors

  • V.A. Manuvera Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 1a Malaya Pirogovskaya str., Moscow, 119435 Russia
  • E.N. Grafskaia Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 1a Malaya Pirogovskaya str., Moscow, 119435 Russia
  • V.N. Lazarev Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 1a Malaya Pirogovskaya str., Moscow, 119435 Russia

DOI:

https://doi.org/10.18097/BMCRM00170

Keywords:

bacterial plasmids; transfection; human cell line

Abstract

Currently, a large number of reagent kits are commercially available for the isolation of highly purified plasmid DNA for subsequent transfection of human cell lines. However, due to high cost and logistical problems, it may be necessary to isolate plasmid DNA using only the simplest reagents and materials. We present one of the possible methods for such DNA isolation, suitable for routine laboratory use. It is based on well-known principles and methods for plasmid DNA purification, has minimal cost, does not require special skills, and is easily scalable. The technique includes the steps of alkaline lysis, purification with silica particles and gel filtration. It was shown that plasmids isolated using the proposed method transfect human embryonic kidney Expi293F cells no less efficiently than plasmids purified using a specialized Qiagen plasmid maxi kit («Qiagen», USA).

References

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Published

2022-09-22

How to Cite

Manuvera, V., Grafskaia, E., & Lazarev, V. (2022). A Simple Technique for Isolating Plasmids from Escherichia Coli for Efficient Chemical Transfection of Human Cell Culture. Biomedical Chemistry: Research and Methods, 5(3), e00170. https://doi.org/10.18097/BMCRM00170

Issue

Section

EXPERIMENTAL RESEARCH