Post on 12-Feb-2020
Procedure Beckman Coulter ProteomeLabTM PA 800
Steps for sample testing using the SDS MW Method
Prepared By: Rafael Hernandez , Jan 2013
Procedure: Logging in • Turning on instrument PA800
– Refer to “Preconditioning SDS procedure”.
• Logging into windows
– Refer to “Preconditioning SDS procedure”
• Logging into the PA800 software (32 karat 8.0 software)
– Refer to “Preconditioning SDS procedure.
– Choose the SDS MW icon
Procedure: Logging into SDS MW
• On the 32 karat software window (see next slide), log in by typing the following: – Username: PA800
– Password: pa800
– Project: SDS MW (select from scroll down menu)
• If you are not in the Direct Control window, do the following: – select from the top menu, Control
– Select “Direct Control”
– Select “View” (see next slide)
Procedure: Turning on and Calibrating PDA Detector
• Turn the lamp on by clicking on the rainbow icon located on the “Direct Control” view.
• Follow the instruction on the lamp control window (see next slide) to turn it on.
• Calibrate detector by clicking on “Calibrate” on the “PDA Detector Parameters”. It will take about 2 minutes for calibration to complete. If calibration fails, make sure that the lamp remains “on” for at least 20 minutes before repeating the calibration.
Check Buffer Trays used during preconditioning and sample testing
• Go to the “Direct Control” View. – Click on “Home”. Trays-rack will move around until it position
itself in buffer vials BI:A1 and BO:A1. – Then, click on “Load” – Open the lid of the instrument, and position the buffer vials as
indicated in the next two slides (layout of buffer trays and vials). • The first slide lay out is for 6 injections or cycles. If planning to do
more than 6 cycles, an additional row of buffer and gel vials is needed for every 6 cycles.
• The second slide lay out is for 24 injections or cycles.
– Close the lid. Ensure that is completely close otherwise the trays will not move to the “home” position (TI:A1 and TO:A1). • It may be necessary to push down the lid and the front cover where
the capillary cartridge is located.
vial H2O
vial Empty
vial Gel
vial NaOH
vial HCl
vial H2O
vial H2O
vial Waste
vial Gel
vial Waste
vial Waste
vial Waste
A B C D E F
Inlet Buffer (BI) Tray on the left Outlet Buffer (BO) Tray on the right
A B C D E F
Layout of the Buffer Trays/Vials during preconditioning and for a CE run of 6 injections (cycles) max
vial H2O
vial H2O
vial Empty
vial Gel
vial NaOH
vial HCl
vial H2O
vial H2O
vial Empty
vial Gel
vial NaOH
vial HCl
vial H2O
vial H2O
vial Empty
vial Gel
vial NaOH
vial HCl
vial H2O
vial H2O
vial Empty
vial Gel
vial NaOH
vial HCl
vial H2O
vial H2O
vial H2O
vial Waste
vial Gel
vial Waste
vial Waste
vial Waste
vial H2O
vial Waste
vial Gel
vial Waste
vial Waste
vial Waste
vial H2O
vial Waste
vial Gel
vial Waste
vial Waste
vial Waste
vial H2O
vial Waste
vial Gel
vial Waste
vial Waste
vial Waste
A B C D E F
Inlet Buffer (BI) Tray on the left Outlet Buffer (BO) Tray on the right
A B C D E F
Layout of the Buffer Trays/Vials during preconditioning and for a CE run of 24 injections (cycles) max
Procedure: Performing Pre-conditioning
• Perform preconditioning before CE sample testing.
– For preconditioning refer to “Preconditioning SDS_procedure”.
Procedure: Performing CE sample testing using the SDS MW method
• Prepare sample(s) as indicated in “SDS-MW Sample Prep Procedure”.
• For a 24 sample injection or cycle, set the sample vials into the “Inlet Sample Tray” as indicated in the next slide from position A1 (sample 1) to C8 (sample24). The Inlet Sample Tray is located in the back of the instrument sample tray. – For loading sample vials into the instrument follow
“Preconditioning SDS procedure” section called “Check Buffer Trays used during preconditioning”.
Sample 8
Sample 16
Sample 24
Sample 7
Sample 15
Sample 23
Sample 6
Sample 14
Sample 22
Sample 5
Sample 13
Sample 21
Sample 4
Sample 12
Sample 20
Sample 3
Sample 11
Sample 19
Sample 2
Sample 10
Sample 18
Sample 1
Sample 9
Sample 17
A B C D E F
Inlet Sample (IS) Tray located on the left buffer/sample compartment
Layout of sample vials for CE sample run
8 7 6 5 4 3 2 1
Procedure: Performing CE sample testing using the SDS MW method
• Write a sequence.
– In the direct control view, go to “file”, then “sequence”, then “open”, then “SDS20.seq”, and click on “Open” (see next three slides).
• The sequence will open, and must be saved as a different file (ex. SDSMW_date_operator initials)
Procedure: Performing CE sample testing using the SDS MW method
• Write a Sequence (cont’d). On the “Sequence” page, the headings of the chart are the following: – “Run #” indicates the sample injection or cycle for
preconditioning.
– In a 24-sample or cycle format, • Run # 1, 8, 15 and 22 are set up for preconditioning the
capillary every six sample injections. Ensure that the “Method” associated with preconditioning is “20cmPreConditioning.met”.
• Run # 2-7, 9-14, 16-21, and 23-28 are set up for samples and SDS-MW analysis. Ensure that the “Method” associated with these injections is “SDS20Separation.met”.
Procedure: Performing CE sample testing using the SDS MW method
• Write a Sequence (cont’d). – The “Sample Inject Inlet” heading indicates the
position of the sample vial for that run. • For example, for “Run # 2”, the nomenclature “SI:A1” means
sample inlet tray, vial position A1. • Make the necessary changes on the chart that will match the
position of each of your sample vials.
– The “Sample Inject Outlet” heading indicates the outlet of the capillary during that cycle. • For example, for “Run # 2”, the nomenclature “BO:C1”
means buffer outlet tray, vial position C1. • Make the necessary changes on the chart that will match the
position of each of your vials.
Procedure: Performing CE sample testing using the SDS MW method
• The next eight slides contain the program graphical screen for each step during the set up of a sequence.
Sequence Page
Procedure: Performing CE sample testing using the SDS MW method
• Once the “Sequence” is set up, instrument is ready for the CE run.
– Go to the sequence page, top menu and select “Control”.
– Under “Control” select “Sequence run”.
– A “Run Sequence” window will appear, then click on “Start”.