For cloning experiments choose from several high efficiency competent cell strains. These E. coli strains are T1 phage resistant and are endA deficient for high quality plasmid preparations. Additionally, all competent cells from NEB are free of animal products.
NEB offers chemically competent Kluyveromyces lactis cells and variants of this strain that have been tailored for specific protein expression needs. These cells are suitable for transformation with any of our linearized pKLAC series expression vectors.
NEB offers several strains with varying levels of expression control, each with T1 phage resistance and extremely high transformation efficiencies.
- Can I store competent cells at -20°C instead of -80°C?
- Does New England Biolabs offer a methyltransferase free strain of Competent E.coli?
- Does New England Biolabs offer strains of Competent E.coli suited for protein expression?
- What are the advantages of New England Biolabs' competent cells?
- What does the growth curve of NEB Turbo cells look like compared to DH5α?
- What is LysY?
- What volume of DNA can be added into competent cells?
- Which kind of transformation tubes should be used?
- What applications are SHuffle® strains useful for?
- Why did Synthetic Biologist Chris Voigt of MIT choose NEB 10-beta for DNA assembly and cloning?
- What formats are competent cells available in?
- Which strain of Competent E. coli should I use for general cloning?
Avoid Common Obstacles in Protein Expression
Read how to avoid common obstacles in protein expression that prevent interactions with cellular machinery.
- Competent Cell Brochure
- Protein Expression & Purification Brochure
- Characteristics of Select E.coli Strains
- Competent Cell Product Comparison
- Competent Cell Selection Guide
- Troubleshooting Transformation Reactions
- Additional E. coli Strain Genotypes
- Chemical Transformation Tips
- Competent E. coli Genome Sequences Tool
- Electroporation Tips
- Genetic Markers
- Making Unmethylated (Dam- Dcm-) DNA
- McrA, McrBC and EcoKI Strain Phenotypes
- Restriction of Foreign DNA by E. coli K-12
- Agrawal, A., Bisharyan, Y., Papoyan, A, Bednenko, J., Cardarelli, J., Yao, M., Clark, T., Berkmen, M., Ke, N., Colussi, P. 2019. Fusion to Tetrahymena thermophila granule lattice protein 1 confers solubility to sexual stage malaria antigens in Escherichia coli. Protein Expr Purif. 153, PubMedID: 30081196, DOI: 10.1016/j.pep.2018.08.001.
- Leith, E.M., O'Dell, W.B., Ke, N., McClung, C., Berkmen, M., Bergonzo, C., Brinson, R.G., Kelman, Z 2019. Characterization of the internal translation initiation region in monoclonal antibodies expressed in Escherichia coli J Biol Chem. 294(48), PubMedID: 31604819, DOI: 10.1074/jbc.RA119.011008
- Reuter, W.H., Masuch, T., Ke, N., Lenon, M., Radzinski, M., Van Loi, V., Ren, G., Riggs, P., Antelmann, H., Reichmann, D., Leichert, L.I., Berkmen, M 2019. Utilizing redox-sensitive GFP fusions to detect in vivo redox changes in a genetically engineered prokaryote Redox Biol. 26, PubMedID: 31450103, DOI: 10.1016/j.redox.2019.101280
- Narayanan, A., Ridilla, M. and Yernool, D.A. 2010. Restrained expression, a method to overproduce toxic membrane proteins by exploiting operatorâ€“repressor interactions Protein Sci. 20 , PubMedID: , DOI:
- Schlegel, S., Klepsch, M., Gialama, D., Wickström, D., Slotboom, D.J. and de Gier, J. 2010. Revolutionizing membrane protein overexpression in bacteria Microb Biotechnol. 3 , PubMedID: , DOI:
- Reddy, P.T., Brinson, R.G., Hoopes, J.T., McClung, C., Ke, N., Kashi, L. 2018. Platform development for expression and purification of stable isotope labeled monoclonal antibodies in Escherichia coli. mAbs MAbs. 10 (7), PubMedID: 30060704, DOI: 10.1080/19420862.2018.1496879
- Wagner, S., Klepsch, M., Schlegel, S., Appel, A., Draheim, R., Tarry, M., Hö, M., van Wijk, K.J., Slotboom, D.J., Persson, J.O. and de Gier, J. 2008. Tuning Escherichia coli for membrane protein overexpression Proc Natl Acad Sci U S A. 105 , PubMedID: , DOI:
- Ren, G., Ke, N. and Berkmen, M. 2016. Use of the Shuffle Strains in Production of Proteins. Curr Protoc Protein Sci. Aug 1, PubMedID: 27479507 , DOI: 10.1002/cpps.11.
- Hibender, S. Landeta, C., Berkmen, M., Beckwith, J., Boyd, D. 2017. Aeropyrum pernix membrane protein VKOR promotes protein disulfide bond formation in two subcellular compartments. Microbiology. 163 (12), PubMedID: 291309344, DOI:
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Use this protocol for maximum transformation efficiency.
Follow these tips to get superior results.
NEB has a long history in recombinant protein expression and has developed a wide array of solutions for proteins that are difficult to express.
In this video, see how transformation plating can be fast and easy using glass beads.