Yaamini’s Notebook: SRM Assay Day 3

More borrowed PRTC = All samples prepared

The assay is still going well! Emma brought down some more QC and PRTC for me to use. I added 30 µL of the final acetonitrile solvent to the 10 µL of PRTC + BSA, then added that to the mass spetrometer vial. I will need to replace the QC after about twelve more QC injections. I also added 30 µL more acetonitrile solution into the blank vial. I then diluted the 20 µL of stock PRTC from Emma with 30 µL of the final acetonitrile solvent to make 50 µL of a 0.2 pmol/µL PRTC solution, following this protocol. I then prepared the remainder of my samples and placed them on the second plate of the mass spectrometer. I had to remake O66 because when I tried pipetting 15 µL into the sample vial, there was not enough solution.

Table 1. Sample order on second plate for the first injection.

Plate 2 1 2 3 4 5
B O32 O60 O101 O91 O100
C O137 O96 O46 O90 O147
D O30 O31 O131 O35 O24
E O43 O40 O26 O78 O124
F O64 O140 O66 O121 OBLNK2

I took the samples that were on Plate 1 and placed them in the freezer. I then added the order for the Plate 2 samples to the sequence file. Since I have to go to Manchester tomorrow, Emma agreed to help monitor the mass spectrometer. I sent her the plate arrangements for the second injection, and asked her to fun the samples in the same order they were on the plate.

Table 2. Sample order on first plate for the second injection.

Plate 1 1 2 3 4 5
B O17 O56 O106 O10 O51
C O145 O71 O49 O06 O21
D O22 O12 O52 O122 O39
E O128 O103 O14 O01 O118
F O113 O04 O102 O99 O08

Table 3. Sample order on second plate for the second injection.

Plate 2 1 2 3 4 5
B O137 O30 O40 O32 O96
C O46 O100 O64 O147 O90
D O43 O78 O91 O26 O131
E O31 O35 O140 O121 O60
F O66 O101 O124 O24 OBLNK2

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