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Step Number
Description
Part Number
1
1 mL Cryobank vials (96 tubes per rack)
374088
2
8 channel handheld screw cap capper/decapper with stand for
Thermo Scientific Nunc-style tubes
4105NUN
3
Thermo Scientific
™
Matrix
™
96 deep-well automation reservoir
1064-15-6
4
Filter plate
Chromsystems
2–8
Thermo Scientific Versette system
- Base unit
- 96 and 384 channel pipetting module
- 6 position stage
- 96 channel air disp. pipetting head, volume 5–300 µL
650-01-BS
650-02-NTC
650-03-SPS
650-06-96300
4–7
Tips – 300 µL extended-length D.A.R.T.’S
5536
8
Titer plate shaker
4625-1CEQ
9
Thermo Scientific
™
Heraeus
™
Labofuge
™
400 with microplate rotor package
75008371
10
Nunc pierceable 96 well cap mats
Nunc polypropylene microplate - 96 well (1.0 mL)
276002
260252
Table 1. Parts and consumables required for specific steps in the liquid handling procedure
Figure 2. Liquid handling procedure
1. The LIMS produces worklist, and
Cryobank vials containing samples
are organized in a 96 well rack (Figure 3).
2. Vials are decapped (8 channel
decapper) and placed on
Versette system (Figures 4 and 5).
3. Reagent reserviors are filled (IS and
precipitation reagent).
4. The empty filter plate and filled reagent
reservoirs are placed on the Versette
system. D.A.R.T.’S tips are replaced.
5. 100 µL sample is transferred to filter
plate. D.A.R.T.’S tips are replaced.
6. 200 µL IS is transferred to filter plate.
D.A.R.T.’S tips are replaced.
7. 25 µL IS precipitation reagent is
transferred to filter plate.
D.A.R.T.’S tips are discarded.
8. The filter plate is removed, covered and
mixed (plate shaker, 600 rpm, 10 min).
9. The filter plate is placed on top of
the microtitre collection plate and
centrifuged (2,000
g
, 4 min).
10. The collection plate is sealed and
placed on the autosampler for analysis
with the TurboFlowmethod and
LC-MS/MS (Figure 6).
4 minutes
Preparation of two
filter plates prior to
simultaneous mixing
and centrifugation
increases throughput.
Figure 3. Cryobank vials
Figure 5. Versette liquid handling system and
one rack of D.A.R.T.’S tips
Figure 4. 8 Channel
handheld screw cap
capper/decapper
Figure 6. Transcend system and TSQ Vantage mass spectrometer