Zymo research
Mix & Go! E.coli Transformation Kit and Buffer Set
T3002
Bacterial transformation is a basic and essential step in molecular biology. Some of the traditional methods of preparing the E. coli competent cells are time-consuming and less efficient. However, this particular product from Zymo offers a simple method of competent preparation. Here I am describing the approach I use in my lab to prepare competent cells using this kit.
E.coli transformation/competent cells
Freshly grown E. coli culture and Zymo buffers
Grow E. coli culture at low temperature (~22 °C)
Freshly grown E. coli culture in zymo broth/SOB media was centrifuged. Pelleted cells were washed with Zymo buffers according to the protocol provided. Finally, cells were suspended in 3-4 ml of competent buffer, aliquoted and stored. For transformation, less than 1 µg of plasmid was added to a vial of the competent cells, mixed gently and incubated on ice for 10-15 min. Heat shock is not required at this step. Further, 100 µl of the mixture was plated. The efficiency was >10^7/µg.
N/A
Very quick and easy to use. High efficiency.
Recovery step is required for kanamycin resistant plasmids
Excellent kit for efficient transformation. Quick and easy to use. Highly recommended.