- Everything should be prepared as described in "Preparing the HPLC setup"

- Create a folder for the experiment in BsXCube.

- Use remote desktop to wbm29sec to control the HPLC system.

If  you are going to intject up to 100 µL:

- Program the HPLC system using the “quick batch” feature: In "setting", set the storage location for the UV data to the folder created in 3.10. Ensure to activate automatic ASCII conversion of the UV data, storing the ASCII file to the same folder. The method file should be saved locally, to C:\XXX. Choose injection volume and well position. Add the measurement to the queue by pressing the “Start” bottom. The software will ask you to save the batch. Prepare for saving but do not press the “Save” bottom, as this automatically starts the batch.

- Set up data collection parameters in the beamline control system. Your local contact will advise you on the time per frame (usually one second). Depending on the column and flow rate choose the number of frames so that total data collection time is in slight excess to the end time of the SEC run.

- Open the safety shutter and start SAXS data collection. Verify that data are correctly acquired by comparing the newly collected data to the buffer data collected previously and checking that the number of counts in the summed intensity remains constant and matches the expected value. Only if all values are correct start the SEC run by pressing the “Save” bottom for the batch table in the HPLC control software and note frame number as displayed in the camserver software when injection is completed and UV data acquisition commences.

 

If  you are going to inject more than 100 µL:

- Program the HPLC system using the “quick batch” feature: In "setting", set the storage location for the UV data to the folder created in 3.10. Ensure to activate automatic ASCII conversion of the UV data, storing the ASCII file to the same folder. The method file should be saved locally, to C:\XXX. Choose injection volume and well position of the buffer vial. Press the “Start” bottom to save the batch and start acquisition of UV data.

- Set up data collection parameters in the beamline control system. Your local contact will advise you on the time per frame (usually one second). Depending on the column and flow rate choose the number of frames so that total data collection time is in slight excess to the end time of the SEC run.

- Open the safety shutter and start SAXS data collection. Verify that data are correctly acquired by comparing the newly collected data to the buffer data collected previously and checking that the number of counts in the summed intensity remains constant  and matches the expected value. Only if all values are correct inject the sample by turing the external valve with the command "hplc_inject" in the spec OH session.