I just found out that Library Efficiency DB3.1 and One Shot ccdB Survival T1R cells have been discontinued. Do you offer an alternative for propagating ccdB-containing plasmids?
We offer One Shot ccdB Survival 2 T1R cells (Cat. No. A10460) for propagating ccdB-containing plasmids.
What kind of competent cells should I use to propagate Donor vectors and Destination vectors? What cells should I use after the BP or LR recombination reaction?
All Donor vectors and Destination vectors contain the ccdB cell death gene to reduce background of non-recombined BP/LR plasmids. Therefore, growing non-recombined vector requires special cells (One Shot ccdB Survival 2 T1R Competent Cells) which are resistant to the lethal effects of ccdB. On the other hand, general E. coli cloning strains including TOP10 or DH5a may be used for plating the BP or LR reaction, or for propagation and maintenance of recombined Gateway constructs.