Generally, we use E.coli TOP10 strain for most of our gene synthesis services. Whilst for special cases, we may apply Stbl3, EPI400, EPI300 or GenScript proprietary strain like Gentoxic V2, GenITR et.al. The information of what competent cell type used in the service can be shared with our customer (will be shown in the COA file) along with the corresponding bacstab or glycerol stock. Growth conditions to handle different cell type:
TOP10, growth in LB overnight at 37°C
Stbl3, growth in LB for 20~24 h at 30°C
EPI400/EPI300, the pr+B1otocol is as below.
a) Add 4 ml LB media into each test tube. Inoculate each tube with bacterial culture with antibiotic at the proper concentration.
b) Incubate the tubes at 37°C, shaking overnight.
c) Dilute the starting culture (from step b) 1:10 into antibiotic supplemented fresh media.
d) Supplement induction solution with a ratio of 1:1000, grow the culture at 37°C for 4 h with vigorous shaking (approx. 250 rpm).
e) Isolate DNA from the induced culture cells as per protocol provided.
