The kit contains reagents to allow you to perform a bacterial transformation with a plasmid that you provide.
- 20mL Bacterial Transformation Buffer (25mM CaCl2 and 20% PEG 8000)
- 1.5mL of LB broth for outgrowth
- Inoculation loops
- Microcentrifuge tubes
- Sterile Water
- Transfer pipettes
- E. coli DH5a
- E. coli BL21(DE3)
How to transform bacteria using this kit
- The day before your transformation streak out on an LB agar plate with bacteria from a stab and grow at 37C overnight(12-18 hours)
- Pipette 50uL of transformation mix into a microcentrifuge tube
- Using an inoculation loop scrape enough bacteria off the plate to fill the loop and twirl into tube containing 50uL transformation mix
- Using a pipette, gently pipette up and own to break up any clumps of bacteria
- Add the plasmid you would like to transform to the tube containing the bacteria
- Put the tube with bacteria in the fridge or at 4C for 15 minutes
- Put the tube into 42C water for 20-30 seconds. Be careful not to go longer as it will kill the bacteria
- Put your tube back in the fridge or at room temperature for 5 minutes. Dissolve the LB in a tube while you wait
- Add 250uL of LB to your bacteria and plasmid tube
- Incubate at 37C(shaking if possible) for 1-3 hours(increased length increases success) or at room temperature for 4-6 hours
- Plate all 250uL on LB agar plate with the appropriate selection antibiotics
- Grow the plate 12-18 at 37C or 24-30 hours at room temperature.
a great educational tool well informative
Honestly I just needed the DH5a, but after looking at the prices of everything, I saw I could get this with two different types of e coli and transformation reagents for not much more than just the single stab, so it seemed a good value.
This kit will save me a lot of time (and expense) and I know that I can trust the quality of sourcing and aseptic technique used in packaging and preparation of reagents supplied by The ODIN. I'm very happy that both common strains of E. Coli (DH5alpha, for plasmid replication) and BL21 (for protein expression) are included with the kit.