Generally, we use E.coli TOP10 strain for most of our gene synthesis services. However, we may use Stbl3, EPI400 or EPI300 strains for special cases. The information of competent cell type used in the service can be shared with our customer along with the corresponding bacstab or glycerol stock. The following is the growth conditions for the different cell types:
1. TOP10, growth in LB overnight at 37°C
2. Stbl3, growth in LB for 20 to 24 h at 30°C
3. EPI400/EPI300, the protocol is as below:
a) Add 4 ml LB media supplemented with antibiotic at the proper concentration into each test tube. Inoculate each tube with bacterial culture.
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.
