Assay

1. Anesthetized 4 mussels in 1 L of 50 g/L MgCl water (600 mL fresh, 400 saltwater).
2. Numbered them with wire tags (8,9,10,12)
3. Treatments (~10 am that day) 8: Given the scrambled negative control from IDT (100 uL, 1 mol). Injected in tip of foot. 9: Injected with 100 uL of PBS in tip of foot. 10: No injection 12: Injected with the 626 (mgfp-3A) siRNA in the tip of the foot (100 uL, 2 mol, 45.205 ug total)
4. Allowed to sit and recover for 33 hours in tub of seawater in seatable with airstone.
5. Harvested at ~7 pm the next day. Mantle and foot tissue were taken (mantle served as control, uninjected tissue).
6. Tube ID’s and treatments stored here.
7. Will do qPCR in Seattle to determine if knockdown actually worked. TBD.

Side note: mussels did not lay threads post recovery. It’s been hard to get them to lay threads all summer, might be a food or stress issue. Also, I let it incubate for ~36 hours because I’ve read that siRNA’s need around 24-48 hours to incubate before testing. People have also tested 7 days after.

Quote of the day

“I am cordially inviting you to grow up”