Submitted 4uL (108ng/uL) of mRNA from gigas gill (from DATE!; sometime in late August) to the Center for Array Technologies (CART) at UW. Sample was labelled “SR01″.
Results:
Results were received on 20090210. Comment from CAT staff member was that it looked “pretty good.” Here are links to the gel (look for SR01), the electoropherogram of SR01, and the EGRAM of the ladder.
Samples were pelleted and washed with 70% EtOH according to Ambion PolyA Purist protocol. Pellets were resuspended in 10uL of The RNA Storage Solution (included in the Ambion PolyA Purist Kit).
Results: The gill mRNA looks great! Good yield and good ratios. Hemocyte mRNA looks kinda rough and a very low yield (which was to be expected).
mRNA was isolated according to Ambion PolyA Purist protocol. After mixing samples with resin, samples were incubated @ RT for 1hr. Samples were washed per the protocol. However, the hemo sample was not clearing from the spin columns with the protocol-directed 3 min. spins. The column had to be spun up to 15 mins. in order for the column to clear. 
Results: The gill mRNA looks good (~1.8ug). The concentration of the hemo sample is extremely low and is below the error threshold for the NanoDrop, but it may not be that bad. The samples are in a relatively large volume (~200uL) and the yield is expected to be very small. So, I will precipitate these samples O/N @ -20C according to Ambion PolyA Purist protocol in order to concentrate them.
The two RNA samples from yesterday were precipitated and washed according to the Ambion PolyA Purist protocol and resuspended in 50uL of 0.1% DEPC-H2O.
Results: RNA readings look better than they did prior to precipitation. The hemo RNA samples will be combined with previous hemo RNA samples and mRNA will be isolated using the Ambion PolyA Purist Kit.
