Isolated RNA from 40 Tanner crab hemolymph samples selected by Grace with the RNeasy Plus Micro Kit (Qiagen) according to the manufacturer’s protocol, with the following modifications:
- Added mercaptoethanol (2-ME) to Buffer RLT Plus.
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All spins were at 21,130g
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Did not add RNA carrier
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Used QIAshredder columns to aid in homogenization and removal of insoluble material
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Eluted with 14uL
RNA was quantified using the Qubit RNA HS (high sensitivity) Assay and run on the Roberts Lab Qubit 3.0.
Used 1uL of sample for quantification.
RNA was returned to the -80C box from where original samples had been stored (Rack 2, Row 3, Column 4).
RESULTS
Qubit quantification (Google Sheet):
Overall, the results aren’t great. Only 15 samples (out of 40) had detectable amounts of RNA. Yields from those 15 samples ranged from 40ng – 300ng, with most landing between 50 – 100ng.
Will pass info along to Grace. Will likely meet with her and Steven to discuss plan on how to move forward.