Geoduck hemolymph calcium assay
made new 20uL 1:1 dilutions from hemolymph samlpes in 1x Calcium Assay Buffer and trashed the old dilutions since
protocol didn’t mention storing dilutions was possible realiquoted 20uL gonad supernatent and didn’t dilute
used the standards diluted on Friday and stored at 4C over the weekend
loaded 10uL each sample into columns 1-4 of the 96-well clear bottom assay plate according to the
plate map under ‘PlateMap’ tab on this googlesheet Mixed Calcium Assay reagent 1 and 2 in a 1:1 mixture (made 10 mL total) and used a reservoir and a multi-channel pipette to add 200uL to all wells. Wells tunred purple!
met Emma at Genome Sciences to read the plate at 575nm
samples were much higher than the standards so I went back and diluted them further making 1:12 dilutions of hemolymph samples by adding 50uL 1x Calcium Assay Buffer to the remaining 10uL of 1:1 diluted samples, and 1:1 dilutions of gonad samples since these were not too far off
loaded 10uL each sample and standards into columns 5-8 of the 96-well clear bottom assay plate according to the
plate map under ‘PlateMap’ tab on this googlesheet re-read the plate again,
data here made some plots in the
xls files to get a rough idea if Ca might correlate with reproductive development, but it wasn’t obvious. I will do a more thorough look later.