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Gonad Fixation


Reagents :

  • 0.1% Tween PBS
  • 3% paraformaldehyde
  • Methanol (Stored at -20°C)

Materials:


Procedure:

  1. With the drawn pipette, remove as much of the solution as possible. Be careful not to disturb the dissected worms.
  2. Add 2 mL of 3% paraformaldehyde (PF) down the side of the dissection dish. Cover with parafilm, and leave in the dark for EXACTLY 1 HOUR.
  3. Add 1 mL of 0.1% Tween PBS down the side of the dissection dish. Swirl the dish gently to get the gonads in the middle of the plate.
  4. With a drawn pipette, gently collect the worms into 2 eppendorf tubes.
  5. Spin gently 1000 rpm for 1 minute in centrifuge.
  6. Wash with 0.01% Tween PBS.
  7. Add 1 mL of MeOH. Place in -20°C for 5 minutes.
  8. Wash 3x with 0.1% Tween PBS.
  9. If necessary, proceed to Step 6 in the DAPI staining protocol.

 

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