Commands and Tips for Thermo Scientific Devices
Finnigan/TSP AS3500/AS3000 Autosamplers: Sample Preparation

Follow these guidelines to ensure successful sample preparation with an AS3500 or AS3000 Autosampler:

• Place extra vials in the three positions after the sample vial position (FirstVial+1, FirstVial+2, and FirstVial+3).

• When you create the Program, make sure the sample prep steps are in the correct order and that all required steps are present. For guidance, refer to the AS3500/AS3000 Basic Template example program shown below.

• Make sure that each dilution cycle includes all of the required steps.

• Specify the desired final volume in the Add step.

• To skip any step (except the Add step), set its volume to 0.

• To perform an operation on the target vial (but not deliver sample to it), set the Add step’s volume to 0. This is useful, for example, when you want to mix or heat the contents of a vial.

• Make sure the volume specified in the Add step in a given dilution cycle is less than or equal to the sum of the PickUpLg and the Load volumes.

• Remember that the autosampler always injects from the FirstVial, which is specified in the Pos. column of the sequence.

Example AS3500/AS3000 Sample Preparation Programs

1. AS3500/AS3000 Basic Template

; Basic Template for sample preparation

; Injection will be made from FirstVial

; Optionally specify Reagent positions

ReagentA = A35

ReagentB = B35

ReagentC = C35

; Required to tell autosampler prep is starting. Must be first command!

PrepStart ; required

; Prep steps defined. Up to 4 sets. Must be done in this exact order!

PrepLoad Volume=0.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=0.0, Reservoir=FirstVial+1 ; optional

PrepPickUp2 Volume=0.0, Reservoir=FirstVial+2 ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial+3 ; optional

PrepAdd Volume=0.0, Reservoir=FirstVial ; required

PrepMix PreHeatMix=0.00, Heat=0.00, PostHeatMix=0.00
 ; optional

PrepWait Time = 0.00 ; optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

;Repeat previous commands for additional 3 cycles

;...

; required end of prep steps. Must be last commands!

PrepFlushP Volume=0.0, Solvent=Solvent_1 ; optional

PrepEnd ; required

0.000 Inject ; Required for sample prep to execute.

End

2. AS3500/AS3000 Dilution Template

; Template is a dilution program that you can use to perform a 1:5 dilution

; Injection will be made from FirstVial

; Required to tell sampler prep is starting Must be first command!

PrepStart ; required

; Prep steps defined. Up to 4 sets. Must be done in this exact order!

PrepLoad Volume=500.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=100.0, Reservoir=FirstVial+1 ; optional

PrepPickUp2 Volume=0.0, Reservoir=FirstVial+1 ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial+1 ; optional

PrepAdd Volume=500, Reservoir=FirstVial ; required

PrepMix PreHeatMix=0.50, Heat=0.00, PostHeatMix=0.00
 ; optional

PrepWait Time = 0.00 ; optional

PrepRemove Volume=0.0, Height=0.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; required end of prep steps. Must be last commands!

PrepFlushP Volume=0.0, Solvent=Solvent_1 ; optional

PrepEnd ; required

0.000 Inject ; Required for sample prep to execute.

End

3. AS3500/AS3000 DABS-CL Template

; Template heats your samples to a given temperature

; Injection will be made from FirstVial

; Turn on heater and set to desired temperature for mix/heat step

Heater =  On

HeaterTemperature = 65

; Optionally specify Reagent positions

ReagentA = A35

ReagentB = B35

ReagentC = C35

; Required to tell autosampler prep is starting. Must be first command!

PrepStart ; required

; Prep steps defined. Up to 4 sets. Must be done in this exact order!

; Cycle 1

PrepLoad Volume=200.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=100.0, Reservoir=ReagentA ; optional

PrepPickUp2 Volume=40.0, Reservoir=ReagentB ; optional

PrepPickUpLg Volume=0.0, Reservoir=ReagentC ; optional

PrepAdd Volume=140, Reservoir=FirstVial ; required

PrepMix PreHeatMix=0.30, Heat=4.50, PostHeatMix=0.30
 ;optional

PrepWait Time = 0.00 ; optional

PrepRemove Volume=0.0, Height=0.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; Cycle 2

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=0.0, Reservoir=ReagentA ; optional

PrepPickUp2 Volume=0.0, Reservoir=ReagentB ; optional

PrepPickUpLg Volume=0.0, Reservoir=ReagentC ; optional

PrepAdd Volume=860, Reservoir=FirstVial+1 ; required

PrepMix PreHeatMix=0.30, Heat=0.00, PostHeatMix=0.00
 ; optional

PrepWait Time = 0.00 ;optional

PrepRemove Volume=0.0, Height=0.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; required end of prep steps. Must be last commands!

PrepFlushP Volume=0.0, Solvent=Solvent_1 ; optional

PrepEnd ; required

0.000 Inject ; Required for sample prep to execute.

End

4. AS3500/AS3000 Rainbow Template

; Prepares four samples

; Injection will be made from FirstVial

; Setup Reagent positions

ReagentA = A35

ReagentB = B35

; Required to tell autosampler prep is starting. Must be first prep command!

PrepStart ; required

; Prep steps defined. Up to 4 sets. Must be done in this exact order!

; Cycle 1

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=30.0, Reservoir=Reagent_B ; optional

PrepPickUp2 Volume=0.0, Reservoir=Reagent_A ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial ; required

PrepMix PreHeatMix=0.50, Heat=0.00, PostHeatMix=0.00
 ; optional

PrepWait 0 ; optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; Cycle 2

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=20.0, Reservoir=Reagent_B ; optional

PrepPickUp2 Volume=10.0, Reservoir=Reagent_A ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial+1
 ; required

PrepMix PreHeatMix=0.50, Heat=0.00, PostHeatMix=0.00
 ;optional

PrepWait 0 ;optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; Cycle 3

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=10.0, Reservoir=Reagent_B ; optional

PrepPickUp2 Volume=20.0, Reservoir=Reagent_A ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial+2 
 ; required

PrepMix PreHeatMix=0.50, Heat=0.00, PostHeatMix=0.00 
 ;optional

PrepWait 0 ;optional

PrepRemove Volume=0.0, Height=2.0 ;o ptional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; Cycle 4

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ;optional

PrepPickUp1 Volume=0.0, Reservoir=Reagent_B ; optional

PrepPickUp2 Volume=30.0, Reservoir=Reagent_A ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial+3
 ; required

PrepMix PreHeatMix=0.50, Heat=0.00, PostHeatMix=0.00 
 ; optional

PrepWait 0 ;optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; required end of prep steps. Must be last of the prep commands!

PrepFlushP Volume=0.0, Solvent=Solvent_1 ; optional

PrepEnd ; required

0.000 Inject ; Required for sample prep to execute.

End

5. AS3500/AS3000 Linear Template

; Template allows you to do linear dilutions

; Injection will be made from FirstVial

; Required to tell autosampler prep is starting. Must be first prep command!

PrepStart ; required

; Prep steps defined. Up to 4 sets. Must be done in this exact order!

; Cycle 1

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=100.0, Reservoir=FirstVial+3 ; optional

PrepPickUp2 Volume=0.0, Reservoir=FirstVial ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial+2 ; required

PrepMix PreHeatMix=0.30, Heat=0.00, PostHeatMix=0.00 
 ;optional

PrepWait 0 ; optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; Cycle 2

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=50.0, Reservoir=FirstVial+3 ; optional

PrepPickUp2 Volume=0.0, Reservoir=FirstVial ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial+1 ; required

PrepMix PreHeatMix=0.30, Heat=0.00, PostHeatMix=0.00
 ;optional

PrepWait 0 ; optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; Cycle 3

PrepLoad Volume=1000.0, Reservoir=Solvent_1 ; optional

PrepPickUp1 Volume=5.0, Reservoir=FirstVial+3 ; optional

PrepPickUp2 Volume=0.0, Reservoir=FirstVial ; optional

PrepPickUpLg Volume=0.0, Reservoir=FirstVial ; optional

PrepAdd Volume=1000.0, Reservoir=FirstVial ; required

PrepMix PreHeatMix=0.30, Heat=0.00, PostHeatMix=0.00
 ;optional

PrepWait 0 ;optional

PrepRemove Volume=0.0, Height=2.0 ; optional

PrepRetvial ; required

PrepMovearm ; required

PrepFlush ; required

PrepFlushBtl ; required

; required end of prep steps. Must be last of the prep commands!

PrepFlushP Volume=0.0, Solvent=Solvent_1 ; optional

PrepEnd ; required

0.000 Inject ; Required for sample prep to execute.

End