What are the best ways to prepare good competent BJ5183 cells?
The most critical step for using the AdEasy system is to make good competent BJ5183 cells. These cells tend to be less efficient than what for transformation. To make good competent BJ5183, you should: 1). eliminate any contaminating strains by growing the cells in the presence of streptomycin; 2). chill the cells longer on ice (2-4hours); 3). thoroughly wash the cells with 10% glycerol (2-3 times); and most importantly, keep the cells concentrated (e.g. one liter culture yields <2 ml competent cells).
