- NF_Tank2_160 (224) -> 10,500 total: 500 for DNA, rest to NF_SetB
- NF_Tank2_160 (100) -> swimmers only
- HC_Tank2_160 (224) -> 9,187 total: 500 for DNA, 5,187 added to HC_SetB, rest to cultch set
- HC_Tank2_160 (100) -> swimmers only
- SS_Tank2_160 (224) -> 11,900 total, added to SS cultch
- SS_Tank2_160 (100) -> swimmers only
- Did live/dead counts for both A and B
- Rinsed tiles (have been doing this everyday I’m in)
- New totals
- NF_SetB: 45,137
- HC_SetB: 60,000
- no new larvae
- Cleaned broodstock buckets
- Rinsed cultch and tiles
Extracted DNA from 24 larvae samples that would be good candidates for test 2b-RAD libraries. I chose sets of new larvae (“LC” for larvae catch), “160”s, and “224”‘s with 3-5 days in between.
|Population||Tank||Family||Size||Date||Storage||Est. #||Date extracted|
|1||Hood Canal||LC||>100||7/13/2015||75/95 EtOH||8/11/2015|
|2||Fidalgo Bay||NA||LC||100||7/13/2015||75% EtOH||8/11/2015|
I still have to go back over my notes to get estimated number for some of them. Storage was mostly in RNALater. A lot of the samples had some white precipitate on the bottom. If larvae weren’t in the precipitate I sucked it out before adding the lysis buffer/proteinase K. Also had the same issue previously with larvae being buoyant in the RNALater even after a spindown. Halfway through trying to siphon off as much liquid as I could, I did some research and found that addition of some ice-cold PBS will change the density of the liquid and allow the larvae to settle out. This worked really well and was done for 2,10,13,15,19,20,21,24. It also dissolved most of the white precipitate.
Followed the protocol I listed here, with a 2.5 hour digest. Could not findd the gel rig set-up (found out later it’s in a different building).