StepOne™ and StepOnePlus™ Real-Time PCR Systems Support – Troubleshooting

Check that all the cables are correctly plugged into the respective ports and not bent. Follow the directions on page 135 of the StepOne™ and StepOnePlus™ Maintenance Guide to check the connectivity. If the issue is still not resolved, please contact Instrument Services at instrumentservices@thermofisher.com for further assistance.

Please send the file to techsupport@thermofisher.com so that we can have our software group attempt to recover the data.

This error means that the data collection was not turned on as appropriate. As shown below, typically the data collection is turned on below the annealing/elongation step. If this is inadvertently turned off, no data will be collected. Thus, the data are not retrievable in such a file.

This issue can occur if duplicate samples are added to the experiment, if illegal characters are used in the experiment/file name, or if the file extension was changed to .eds from a different extension.

Please send the file to techsupport@thermofisher.com to attempt recovery.

This issue can occur if the “Start Run” button is clicked more than once or if illegal characters are used in the experiment/file name. You can recover the data yourself by overriding the calibration file with one that represents the calibration status of the instrument when you ran that particular file. Use the toolbar at the top to navigate: Analysis → Override Calibration → Use Calibration From Another File.

This is generally caused by incorrect placement of tube strips, resulting in a sample block that can’t properly be lowered. We recommend contacting Instrument Services (1-800-955-6288, option 3, option 1), to have an engineer walk you through a manual drawer-lowering process during instrument power-up. To prevent this from happening in the future, place tubes toward the middle of the block or use a placeholder strip on the opposing side to counterbalance block pressure.

This issue can occur if the run is interrupted. To prevent this from happening in the future, avoid ending the PCR with an infinite hold stage or interrupting the run. Please send the file to techsupport@lifetech.com so that our software group can attempt to recover the data.

This can occur if the tubes are not being used with the VeriFlex™ tray. The tubes are actually being crushed, not melting. Please follow these instructions for proper loading of tubes and tube strips.

• Put the tubes or tube strips in the tray with the adapter on the 96-well support base.
• If using a plate, put the plate directly on the 96-well support base.
• Pipet your reactions into the tubes/tube strips or the wells on the plate.
• Seal the tube/tube strips with optical flat caps.
• If using a plate, seal the plate with either optical adhesive film or flat caps.

For optimal performance on the StepOnePlus™ instrument with partial loads:

• Load at least 16 tubes and arrange them in adjacent columns of 8 tubes, using rows A through H, or adjacent rows of 8 tubes, using columns 3 through 10.

For optimal performance on the StepOne™ instrument, load at least 4 tubes in the sample block.