e5 sample prep for lipid and metabolite analysis

To characterize lipids and metabolites in the e5 timeseries data, we have to homogenize and separate the samples before sending them to the UW facility. We first sent them 4 test samples to see if these samples would work. The lipids worked great but the metabolite data was lacking some key metabolites – we may not have given them enough sample as input and the instruments may have gotten disrupted by salt since we used 10x PBS to airbrush the samples. See the former protocol e5 timeseries molecular github for information about test samples.

To remedy these issues, Ariana and I developed the following protocol, which I followed for all samples unless otherwise noted.

The major changes of the protocol:

• Use LCMS grade water for airbrushing
• Airbrush fragment primarily with air and remove any residual tissue with water + airbrush, which should lead to a more concentrated sample
• Collect homogenate from each sample for protein assay
• Run protein assay to quantify the ug/mL of protein in tissue

Sample prep

Below are the details for the samples I prepped. There are 96 samples total. I will continue to add to this table as I complete each sample.

In the table, the serial number is the number on the tube that will be sent to UW for analysis.