The overall goal of this project is to better understand senescence in local sockeye salmon populations. This will lay the groundwork for exploring significant variation observed at the population level. Samples will be taken from at least two time points during their upstream migration. Tissues samples will include brain (and maybe liver) n=20
Differences in gene expression patterns will be measured to better understand physiological changes that are taking place during this life stage.
Protocol:
Cut samples to a maximum thickness of 0.5 cm in any one dimension and submerge in 5 volumes of RNAlater.
Try to not preserve >1 cubic cm (1 cc would require 5 ml or RNAlater).
Make sure tissue is submerge (not on side of tube)
Once in RNAlater, samples can be stored for up to 1 day at 37°C, 1 week at 25°C, 1 month or more at 4°C, and long-term at -20°C or -80°.
Samples in RNAlater can safely be shipped on wet ice for several days. For longer shipping times use dry ice.
Minimum two sites, three tissues @ each site; 10 samples each tissue. Minimally: 60 samples.
Record sex, length, weight, etc.
Try to get close to 1:1 sex ratio. 2:1 is fine, ie minimum of 3 per sex
Shipping 250 ml or RNAlater and 100 tubes. Can easily send more if needed.
RNaLater (250ml): $175
1/2 kit: Brilliant II SYBR Green qPCR MM $219
Tri-Reagent (200ml): $220