Preparing homemade XL1-blue cells in CCMB80 buffer - (Jul/30/2012 )
I have read several topics from this forum which recommend preparing homemade competent cells in CCMB80 buffer instead of CaCl2. This purportedly increases transformation efficiency by several fold or even orders of magnitude. Most topics link to this protocol, (http://openwetware.o...competent_cells) which has been tested on several strains, including Top10 and DH5α.
I would like to know if this method has been tested with XL1-blue cells and if using XL1-blue would have a higher transformation efficiency using this method than DH5α. I am attempting a primer extension mutagenesis based on megaprimer extension (http://www.biotechni...18_O_95037a.pdf), but my transformations haven't produced any colonies yet. I'd like to prepare highly efficient competent cells that can be used to perform transformations of nicked circular DNA. Could I achieve this using the CCMB80 method on XL1-blue cells?
I don't see why not, most methods of making competent cells work in a similar fashion, so it should be fine.