限制性内切酶酶切的常见问题及解决方法（The common problems and solutions of restriction endonuclease digestion）.docVIP

限制性内切酶酶切的常见问题及解决方法（The common problems and solutions of restriction endonuclease digestion） The common problems and solutions of restriction endonuclease digestion Restriction enzyme digestion of the common problems and solutions, personally feel very well summed up, specially posted for you to share. Enzyme digestion problems, see enzyme instructions, the corresponding reagent company directory. The enzymes produced by different companies, the source of strains, the preparation process, purity, viability and enzymatic activity may be optimized differently,. Can be found in the above definition, unit of enzyme preservation conditions, enzyme system of [buffer and other enzymes such as buffer, double cut enzyme reaction temperature [some enzymes in 50, 55 or 30 temperature reaction, whether methylation affects the enzyme activity and whether there is an asterisk, asterisk may appear live factors and protective base, isocaudarner, isoschizomer 1. enzymes can not be cut or incomplete The 1.1 plasmid is the most common or poorly expressed inhibitor. The presence of miscellaneous proteins can affect enzyme digestion, showing A260/A280 below 1.8; inhibitors are common in phenols, salts, and ethanol; [re DNA, using reliable kits or reliable manual extraction reagents, 1.2 enzyme problems: make sure the enzyme is effective [although many enzymes have expired time, but expired as long as the enzyme can be effectively cut, available. Make sure the enzyme is not effective. Mark it and replace it. [note] topic: by endonuclease A. endonuclease, if no special requirement, keep -20~-30 degrees. Is not as low as possible, the enzyme is usually preserved in 50% glycerol buffer, when the temperature is too low, will freeze (enzyme transport process exception, because the enzyme requires low temperature transport, so the convenient situation is the use of dry ice, the enzyme will freeze, but the freezing times is limited, is a relatively better choice), if is often used, the enzyme wi