Laura’s Notebook: SRM Dilution Curve Sample Prep

Prepping dilution curve samples

After we run all our samples Yaamini & I are going to run 10 samples with mixtures of geoduck/oyster protines. We will prepare them at different, known preportions of G:O, and see how the peptides we each targeted become more/less prevalent. This is good practice to prove that we are measuring what we say we are.

Yaamini did a great job calculating/writing up the dilution calculations (after a couple iterations, see here and here

As Yaamini did, I pooled 5ul of 5 randomly selected samples for a total of 25ul of pooled sample – I selected samples that had the full 2ug/ul (as opposed to those that had low concentration), pipetted them into a centrifuge tube, vortexed, then spun down gently:

  • 120
  • 47
  • 79
  • 60
  • 9

Then, I got Yaamini’s dilution curve samples, that already have PRTC & ACN+FA, out of the freezer, and pipetted the designated vol of my pooled geoduck sample into each tube:

image

Then I vortexed each, spun down gently, labeled autsampler vials, and transferred the dilutions into the vials. I put them in the -20 freezer until we are ready to use them.

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