PMID:21362398
| Citation |
Warren, DJ (2011) Preparation of highly efficient electrocompetent Escherichia coli using glycerol/mannitol density step centrifugation. Anal. Biochem. 413:206-7 |
|---|---|
| Abstract |
Traditional protocols for preparing Escherichia coli for electroporation are laborious and often deliver highly variable transformation efficiencies. Many laboratories resort to purchasing expensive commercially prepared cells. This article describes a simple method for producing electrocompetent E. coli by centrifuging bacteria through a glycerol/mannitol density cushion. The method is rapid and replaces tedious multistep procedures with two 15-min centrifugations. Standard cloning strains consistently produce more than 8×10(9)transformants/μg pUC18, whereas the strains TG1 and LE392 display efficiencies of more than 3×10(10)/μg DNA. |
| Links |
PubMed Online version:10.1016/j.ab.2011.02.036 |
| Keywords |
Centrifugation, Density Gradient/methods; Electroporation/methods; Escherichia coli/cytology; Escherichia coli/isolation & purification; Glycerol/chemistry; Mannitol/chemistry |
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