Question (2026)
Question
How do I revive an agar stab?
ABM community
Verified customer
Asked on Mar 24 2025
Answer
1. Use a sterile loop or pipette tip to scrape a small amount of cells from the agar stab. Only a tiny amount is needed as it contains live E. coli.
2. Gently streak the cells onto an agar plate containing the correct antibiotic selection in order to achieve single isolated colonies.
3. Place the plate “agar side up” and incubate at 37°C overnight.
4. Select a single colony from the plate for downstream applications, such as an overnight broth culture and grow E. coli to late exponential-early stationary phase. 4a. The broth culture can be subjected to miniprep plasmid extraction. We recommend using abm’s Column-Pure Plasmid Miniprep Kit (Cat. No. G4003). 4b. The broth culture can be used to prepare a glycerol stock for long-term storage. Mix culture with sterile glycerol to a final concentration of 15% and store at -80°C.
2. Gently streak the cells onto an agar plate containing the correct antibiotic selection in order to achieve single isolated colonies.
3. Place the plate “agar side up” and incubate at 37°C overnight.
4. Select a single colony from the plate for downstream applications, such as an overnight broth culture and grow E. coli to late exponential-early stationary phase. 4a. The broth culture can be subjected to miniprep plasmid extraction. We recommend using abm’s Column-Pure Plasmid Miniprep Kit (Cat. No. G4003). 4b. The broth culture can be used to prepare a glycerol stock for long-term storage. Mix culture with sterile glycerol to a final concentration of 15% and store at -80°C.
ABM Scientific Support
Answered on Mar 24 2025