Mon. Dec 10, 2018

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.

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Written on December 10, 2018