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The FLOW PCR Setup Instrument is intended for general laboratory use and must be used exclusively by professional users trained in laboratory techniques and having studied the user documentation of this instrument.

The FLOW PCR Setup System performs pipetting operations on liquids from secondary tubes and MagNA Pure 96 Output Plates to LightCycler® 480 Multiwell Plates.

Pipetting means transferring small quantities of liquid from one vessel to another. A pipetting operation is achieved by aspirating liquid from a source vessel, then transferring and dispensing it into the target vessel.

The FLOW PCR Setup System is used in combination with the FLOW Solution. The FLOW Solution provides an automated solution for a laboratory workflow comprising primary sample handling, purification, PCR setup and qPCR.

The FLOW PCR Setup Instrument can pipette multiple components to prepare different qPCR reaction mixes. After dispensing the reaction mix into the LightCycler® 480 Multiwell Plate(s), the FLOW PCR Setup Instrument transfers eluates from MagNA Pure 96 Output Plates to the reaction mix in the LightCycler® 480 Multiwell Plate(s).

The FLOW Primary Sample Handling Software can be installed and executed on the FLOW PCR Setup System. This supports a new FLOW Solution configuration, FLOW Reloaded, with primary sample handling and PCR setup enabled on the same system. The FLOW Primary Sample Handling and MagNA Pure 96 System workflow can also be established using a FLOW PCR Setup Instrument in primary sample handling mode.
When using this FLOW Reloaded or FLOW PSH on PSU Standalone configuration, please follow the instructions in the FLOW Primary Sample Handling System Operator's Guide for primary sample handling on the FLOW PCR Setup System. The technical specifications of the FLOW Primary Sample Handling System also apply to the FLOW PCR Setup System when operated in primary sample handling mode.

For PSH workflow on PSU, the following accessories should be ordered for primary sample handling on a PSU instrument:

The following accessories are mandatory to operate the PSU in PSH mode:
04 639 502 001Rack-SMP-CAR-24-A00 (set of 4)
04 639 529 001Rack-SMP-CAR-32-A00 (set of 3)
07 101 970 001 FLOW Processing Cartridge Carrier

The following accessories are part of the PSU delivery, but one additional copy should be ordered in addition to avoid having to use the same carrier for both PSU- and PSH- modee:
04 639 545 001Rack-TIP-CAR-480-A00 for 480 tips
07 127 014 001 FLOW 32 vial carrier with adaptors (set of 3)

Please be aware, that accesories listed above are required to operate the PSU in PSH mode.
This information is not clearly stated in the FLOW PCR Setup System Operators’ Guide V5.0 SW 2.1 and Addendum 1 - May 2020 (page 42).
The information will be complemented accordingly with the next update of the operators guide.
Optional:
07 101 988 001 Primary Sample Handling Reagent Carrier (for PSH Standalone workflow only)
04 639 618 001 Rack-Tube Carrier Filling Rack (optional)

", "Country": "XG", "Code": "Background Information", "Name": "Background Information" }, { "Language": "en", "Value": "The FLOW PCR Setup Instrument (PSU) is a benchtop robotic workstation for fully automated PCR reaction mix setup and distribution into PCR plates from individual components or ready to use reagents. The FLOW PCR Setup Instrument includes automated tracking of barcoded accessories and consumables. The hardware is designed for aspirating and dispensing volumes up to 1 ml.

The FLOW Primary Sample Handling Software can be installed and executed on the FLOW PCR Setup System. This supports a new FLOW Solution configuration, FLOW Flex, with primary sample handling and PCR setup enabled on the same system. The FLOW Primary Sample Handling and MagNA Pure 96 System workflow can also be established using a FLOW PCR Setup Instrument in primary sample handling mode.
When using this FLOW Flex or FLOW PSH on PSU Standalone configuration, please follow the instructions in the FLOW Primary Sample Handling System Operator's Guide for primary sample handling on the FLOW PCR Setup System. The technical specifications of the FLOW Primary Sample Handling System also apply to the FLOW PCR Setup System when operated in primary sample handling mode.
", "Country": "XG", "Code": "Product Description", "Name": "Product Description" }, { "Language": "en", "Value": "The PCR Setup Instrument (Cat. No. 07101996001) is delivered together with these items:
  1. PCR Setup Instrument (07101996001) 1 instrument
  2. FLOW 32 Vial Carrier (Set of 3) (07127014001) 2 carriers with adaptors
  3. Rack-TIP-CAR-480-A00, for 480 tips (04639545001) 2 rack tip carriers
  4. Rack-PLT-CAR-L5AC (04639553001) 2 multiwell plate carriers
  5. FLOW PCR Setup LC480 Multiwell Plate Cooling Block (96 well) (07102046001) 6 cooling blocks (96 well)
  6. FLOW PCR Setup LC480 Multiwell Plate Cooling Block (384 well) (07102054001) 6 cooling blocks (384 well)
  7. FLOW PCR Setup Output Plate Cooling Block (07162367001) 3 cooling blocks
  8. FLOW Orbit Reader Cap (07353049001)
", "Country": "XG", "Code": "Content", "Name": "Content" }, { "Language": "en", "Value": "
FLOW PCR Setup Instrument Software Version 2.1

Restart Function

The restart function is not applicable for FLOW PSU SW V 2.0. The SW needs to be closed and restarted after every run!

The FLOW Software does not create the PSU setup worklist

If the FLOW Software does not create a worklist for starting the setup on the FLOW PCR Setup System (PSU setup worklist), there are two ways to continue:
  1. Delete the old PSU setup worklist, as it may block the creation of a new one.
  2. Use the final worklist generated for the FLOW PCR Setup System (PSU sample worklist) as the PSU setup worklist.

To delete the old PSU setup worklist

1. Open the O:\\PSU\\Worklist directory.
2. Delete the Worklist_Setup.xml file.
3. If present, also delete the Worklist_Samples.xml file.
4. Wait a few minutes and refresh the O: drive. The new PSU setup worklist should be created automatically:

-If a PSU setup worklist was created, verify that the samples in the FLOW worklist have the status PURIFICATION_STARTED and resume the FLOW process.
-If there is still no PSU setup worklist, proceed with section To use the PSU sample worklist as PSU setup worklist, below.


To use the PSU sample worklist as PSU setup worklist

If the samples in the FLOW worklist have the status PURIFICATION_STARTED and there is no PSU setup worklist, the PSU sample worklist can be used by copying and renaming it.
1. Finish the purification step.
2. Wait until the status of the samples in the FLOW worklist has changed to PSU_STARTED.
3. Open the O:\\PSU\\Worklist directory.
4. Copy the Worklist_Samples.xml file.
5. Rename the copy Worklist_Setup.xml.


The FLOW Software does not create the PSU sample worklist

.Keep in mind that the time it takes to create the PSU worklist depends on the complexity of your order.

The following are some of the factors that affect the time it takes to create the worklist:
  • The presence of duplicates.
  • The absence of PCR controls.
  • The number of targets for which you want the samples tested.
  • The number of Light Cycler® 480 Multiwell Plates that have to be produced.
  • The presence of an RT reaction.


The instrument run was aborted

The FLOW PCR Setup Instrument Software creates a backup of all worklists and result files that have undergone the sample transfer step. All worklists and result files receive a date-time stamp to prevent overwriting and to facilitate retrieval.

To use the old PSU worklist

1. On the FLOW PCR Setup Control Unit, open the C:\\FLOWFiles directory, containing backups of worklists and result files.
2. To re-use the old worklist, search for the last file named worklist_YYYMMDD_HHMM.xml.
3. Copy this file to the O:\\PSU\\Worklist directory.
4. Rename the file worklist.xml.
5. Start the FLOW PCR Setup Instrument Software and continue with the instrument run.

The FLOW PCR Setup Instrument has no power

If the process on the FLOW PCR Setup Instrument is interrupted because the instrument has no power or has crashed, it is not possible to process the samples manually, because the FLOW PCR Setup Instrument Software will not create a result file. If necessary, the cause of the crash must be resolved, and then the FLOW PCR Setup System can be restarted.

Normally the FLOW process will continue if the FLOW PCR Setup System is restarted. If the worklist files are already processed, but no result file is generated on the FLOW PCR Setup Instrument, the total process needs to be restarted.

To restart the process

1. Press the main switch on the instrument.
2. Load the old PSU worklist. For detailed information, see section To use the old PSU worklist, above.
If the FLOW PCR Setup System is interrupted while it is processing the samples from the MagNA Pure 96 Output Plate, the FLOW process will be terminated. It is not possible to continue manually.
The outstanding orders and the worklists for the FLOW PCR Setup System must be deleted.

To remove outstanding orders from the FLOW worklist

For detailed information, see section To delete all test requests in the FLOW Software (Reset unread tables), above.

To delete the PSU setup worklist and the PSU worklist

1. Open the O:\\PSU\\Worklist directory.
2. Delete the Worklist_Setup.xml file.
3. Delete the Worklist_Samples.xml file.
4. If the worklists cannot be deleted, the unread tables must be reset in the FLOW Software. For detailed information, see section To delete all test requests in the FLOW Software (Reset unread tables),in the FLOW Solution Operator's Manual.

The FLOW PCR Setup Control Unit has crashed

If the process on the FLOW PCR Setup System is interrupted because the control unit has crashed, please contact your local Roche Customer Support representative.


The FLOW Software cannot receive the PSU result file

If the FLOW Software cannot receive the result file from the FLOW PCR Setup System (PSU result file) because there is no network connection and the network connection cannot be restored, the status of the samples in the FLOW Software will not be updated to PSU_COMPLETE and no LC480 order file will be created. In this case the result file must be transferred manually from the FLOW PCR Setup Control Unit to the correct directory on the FLOW Control Unit. This is done with a USB drive.

1. On the FLOW PCR Setup System Control Unit, open the C:\\FlowFiles directory where the result files are located.
2. Copy the [barcode]_[date]_[time].xml file(s) to a USB drive. Make sure you copy the files with the most recent date.
3. Rename these files [barcode].xml.
4. Copy the result file to the O:\\PSU\\Results directory.
5. The FLOW Software will now process the result file and generate an order file for the LC480 instrument (LC480 order file). The samples listed in the result file will receive the status PSU_COMPLETE in the FLOW worklist.

-> When the status is changed to LC480_Started and you have placed the LightCycler® 480 Multiwell Plate in the LC480 instrument, continue with the FLOW process.

->If the status does not change to PSU_COMPLETE and no LC480 order file is generated, the samples must be processed manually. Proceed with section No order file for the LC480 instrument, in the FLOW Solution Operator's Manual


1.5 mL Tube

Consequences of using the wrong tube type
Scenario 1: 2.0mL tube defined in the software but customer incorrectly uses 1.5mL tubes
- No Liquid Level Detection errors, as the system finds more liquid than expected (the liquid level in 2.0mL tubes is lower than in 1.5mL tubes).
- Liquid Level tracking errors can occur. During pipetting, the system follows the liquid in the tube. Due to the difference in shape, the liquid level in 2.0mL tubes retracts more slowly (µL/mm) than with 1.5mL tubes.Therefore the pipette moves slower than the liquid and will aspirate air, resulting in failed reactions.
- No crash as the bottom of both tubes have exactly the same defined and physical height.

Scenario 2: 1.5mL tube defined in the software but customer incorrectly uses 2.0mL tubes
- Liquid level detection errors, the liquid is much lower than expected and calculated as not enough liquid for the next steps.
- No liquid level tracking errors, the liquid is moving slower than calculated. The tips dive more quickly into the fluid than the fluid is retracting. Reactions will be pipetted as expected.
- No crash as the bottom of both tubes have exactly the same defined and physical height.", "Country": "XG", "Code": "Troubleshooting", "Name": "Troubleshooting" }, { "Language": "en", "Value": "PSU is a pipetting sample handling instrument based on the Hamilton STARlet", "Country": "XG", "Code": "Principle", "Name": "Principle" } ] } } ] }

FLOW PCR Setup Instrument

FLOW PCR Setup Instrument

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