Some tips for preparation of E. coli competent cells

  • Lin Zhao
  • Huan Wang
  • Yukun Jia
  • Xianyu Yang
DOI: https://doi.org/10.53555/bs.v4i8.2164
Keywords: E. coli, Competent cells, Preparation

Abstract

In this study, we modified the protocol of E. coli competent cell (CC) preparation on two aspects. One is shortening centrifuge time to 2 min instead of 15 min (used in the traditional method), by which the exposure time of cell in solutions for CC preparation was shortened, and the chemical or biological damage on cells could be minimized. The other is suspending cells with disposable Pasteurized pipette instead of micropipette tips to diminish physical damage on cells. By this modified protocol, CC average transformation efficiency of two batches is 1.88×108 and 2.07×108, respectively, 10 times higher than that of CC prepared by traditional method.

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Author Biographies

Lin Zhao

College of Animal Science and Technology, Zhejiang Agricultural and Forestry University, 666 Wusu Street, Hangzhou-Lin’an, Zhejiang 311300, China

Huan Wang

College of Animal Science and Technology, Zhejiang Agricultural and Forestry University, 666
Wusu Street, Hangzhou 311300, China

Yukun Jia

College of Animal Science and Technology, Zhejiang Agricultural and Forestry University, 666
Wusu Street, Hangzhou 311300, China

Xianyu Yang

College of Animal Science and Technology, Zhejiang Agricultural and Forestry University, 666
Wusu Street, Hangzhou 311300, China

References

1. Lee SY, Chang HN. Generation of bacteriophage lambda lysogens by electroporation. Biotechniques. 1994, 16(2): 206-208.
2. Sambrook J, Fritsch E, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, NY. pp1.82-1.84
3. Tung WL, Chow KC. A modified medium for efficient electrotransformation of E. coli. Trends Genet. 1995, 11(4): 128-129.
Published
2018-08-30
How to Cite
Zhao, L., Wang, H., Jia, Y., & Yang, X. (2018). Some tips for preparation of E. coli competent cells. IJRDO - JOURNAL OF BIOLOGICAL SCIENCE, 4(8), 18-20. https://doi.org/10.53555/bs.v4i8.2164
Issue
Vol. 4 No. 8 (2018): IJRDO - Journal of Biological Science
Section
Articles

Copyright (c) 2018 IJRDO - Journal of Biological Science (ISSN: 2455-7676)

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