We are struggling in getting recombination for baculovirus production in E.coli BAC10 cells. Is there any protocol to make "good" competent cells from BAC10 bacteria? Have some of you experienced the same problems?
Grow your cells in LB media and then do a couple of CaCl2 washes making sure all the while that the cells, buffer, tips, pipettes and any tubes that you use are chilled. In my experience preparing the competent cells fresh or storing them in -80C and then using the cells have made no difference in the transformation efficiency.