Streaking RNAi Bacteria
- Disposable Pipette Tips
- LB/A Plates
- Appropriate Bacteria
- Wear gloves throughout the entire procedure.
- Using a sterile pipette tip, pick a small amount of the desired bacteria from the RNAi library. Push the pipette tip through foil cover and scrape the tip around the inside walls of the well.(Figure 1)
- Cover the well punctured well with a piece of tape. (Figure 2)
- Lightly streak the bacteria onto a fresh LB/A plate. Be very careful to use a light touch and not slice the agar. NOTE: It is our lab's procedure to use one LB/A plate for multiple lines of RNAi bacteria. The plate is divided into various quadrants and the bacteria is streaked in its appropriate location. (Figure 3)
- Grow overnight in a 37°C incubator.
- Remove the plate from the incubator in the morning and store it at 4°C. These stocks will be good for a month or two, or until they become contaminated.
- Use this plate to maintain the strain. Maintenance of the stock is identical to the Sreaking OP50 procedure, just make sure you use LB/A Plates.
Pick appropriate bacteria from RNAi library. Make sure you go into the correct well!
Cover punctured well with tape.
Use one plate when streaking multiple strains of RNAi bacteria.