SHuffle cells have been engineered from two E. coli lineages: K12 (NEB #C3026) and B (NEB #C3028, C3029 and C3030). For a detailed description of the strains, please refer to the SHuffle Strain Choice Table at the bottom of this page.
Proteins express to various degrees of success in SHuffle-K12 vs SHuffle-B (see table below). We therefore recommend the following general protocol for optimal protein expression in SHuffle cells:
- Test the expression of your protein at both SHuffle lineages (K12 vs B) and select the better expressing version.
- Test expression of protein in various temperatures. Grow cells at 30ºC for 5 hours, split cells, and shift to various temperatures. We suggest growing cells overnight for those shifted to lower temperatures (16ºC to room temperature).
- Should further optimization be necessary, continue optimization of inducer concentration, induction time, and various helper plasmids. Refer to our list of publications using SHuffle for further details in expression conditions.