3. Transformation of the plasmid into BL21(DE3) E. coli
- Take out five 50μl E. coli BL21(DE3) competent cells from the -80℃ fridge and place them on ice.
Wait for
5 minutes until the competent cells unfreeze.
- In each of the five microtubes, add in 3μl 50ng/μl of the plasmids (16S_pUC57, ipaH_pUC57, invA_pUC57,
cagA_pUC57, Cas12a_pET-28a). Set it aside for 25 minutes.
- Place the microtube in a 42℃ water bath and heat shock for 45 seconds.
- Place the tube back on the ice for 2 minutes.
- Add 700μl lysogeny broth (LB) into each of the tubes and incubate at 200 rpm for 60 minutes.
- Centrifuge at 5000 rpm for 2 minutes and coat on LB medium plates.