SRM Dilution Calculations
My prior calculations were incorrect!
As I mentioned previously, Laura and I need to run dilutions of our samples to confirm that as the concentration of protein in our samples decreases, the assay does not detect them as intensely. When I showed Emma my calculations, she suggested making dilutions based on protein concentrations (µg) as opposed to µL of sample added. Here are my final numbers:
Table 1. Dilution calculations
|Vial||Dilution Ratio||µg to Inject||µL to Inject||µg oyster Needed||µg geoduck Needed||µL oyster needed ((µg/µL)x15 µL/1 µg/µL)||µL geoduck needed ((µg/µL)x15 µL/2 µg/µL)||PRTC added||µL ACN||Total Volume (µL)|
For PRTC, our stock concentration volume is 0.5 pmol/µL. I changed the injection volume for the 10:1 and 1:10 dilutions from 2 µL to 1 µL to ensure we were pipetting volumes over 0.5 µL. This made it so that the final volume for the 10:1 injection was 15.75 µL, but Emma said that was okay.
Written on July 20, 2017