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Streaking RNAi Bacteria


Materials:

  • Disposable Pipette Tips
  • Gloves
  • LB/A Plates
  • Appropriate Bacteria

Procedure:

  1. Wear gloves throughout the entire procedure.
  2. 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)
  3. Cover the well punctured well with a piece of tape. (Figure 2)
  4. 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)
  5. Grow overnight in a 37°C incubator.
  6. 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.
  7. 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.

Figures:

Figure 1.

Pick appropriate bacteria from RNAi library. Make sure you go into the correct well!

 

back to procedure

 

Figure 2.

Cover punctured well with tape.

 

back to procedure

 

Figure 3.

Use one plate when streaking multiple strains of RNAi bacteria.

back to procedure

worm

Sudhir Nayak, Ph.D.

Biology Building 126
2000 Pennington Rd.
Ewing, NJ 08628
P) 609-771-2659

 

Dr. Nayak’s Lab
Biology Building 236
P) 609-771-3436
E) nayaklab@tcnj.edu