ERP Micro December 2019

test kit and should be stored in the refrigerator (5 ± 3 °C). Between each use the aluminum cooling block should be returned to the refrigerator. 3) The Assurance® GDS Rotor-Gene® set up and data entry should be completed prior to transferring samples from the resuspension plate into the EHEC Amplification Tubes Tq. 4) Remove the Amplification Tubes from the foil pouch and place them in the frozen gel cooling block. Reseal and return the pouch to the refrigerator. 5) Transfer 30 µL of the sample from resuspension plate well into each Amplification Tube using a multi-channel pipette and filter barrier tips. Firmly press down on each Amplification Tube lid to close. Visually inspect each tube to ensure that the cap is securely sealed. 6) Place Amplification Tubes into the rotary ring in sequential order, beginning with position # and load into the Assurance® GDS Rotor-Gene®. 7) For the 576 kit, place amplification tubes into the aluminum cooling block. Place amplification tubes into Assurance® GDS Rotor-Gene® in sequential order, beginning with position #1. For the 100 test and ATM test kits, use the 36- well Rotor-Disc and locking ring; for the 576 test kit, use the 72-well Rotor-Disc and locking ring. 8) Note: For 576 test kit, after loading amplification tubes in the Rotor-Disc and securing with locking ring, contents should be thoroughly mixed by shaking with a snapping motion. See Application Note FRMMK.2060 for additional details. 9) Start the Rotor-Gene®. Refer to Assurance® GDS User Manual (Cat No. 55342BC) for detailed instructions on operating the Rotor-Gene®. Note: The Assurance® GDS Rotor-Gene® must be started within 20 min after the addition of the samples to the amplification tubes. turning on the power to the computer. Open the Assurance® GDS software on the computer and enter the login information. (b) Open the software to get to a new run file. (c) Enter sample identification numbers: select a well to start and enter the well ID in the box “Sample Name” on the right and press the Enter key. (d) Enter the sample description. (e) Enter the Lot Numbers for the kit used. (f) Select “E. coli O157:H7” from the dropdown list of tests on the left side of the screen. Create a Run File (a) Turn on the power to the Assurance® GDS Rotor-Gene® instrument, followed by

seal with the rotary locking ring. (c) Close the instrument. (d) Select “Start PCR Run”.

Note: Wear new gloves prior to handling reagents. (a) Prior to initial use, cool the gel cooling block, B ( g ), in the freezer (–25 to –15°C) for 6 h. When not in use, the gel cooling block should be stored at –25 to –15°C. Between each use, return the gel cooling block to the freezer until it has turned completely purple, indicating it is ready for use. This may take up to 2 h. (b) Remove amplification tubes, C ( d ), from foil pouch and place the amplification tubes in the gel cooling block. Reseal pouch. (c) Open caps of amplification tubes in gel cooling block. Transfer 30 µL of sample from the resuspension plate wells into each amplification tube using the eight-channel pipettor and filter- barrier tip, B ( j ). Firmly press down on each amplification tube cap

Loading the Assurance® GDS Rotor-Gene® (a) Open the instrument lid. (b) Place the sealed Amplification tubes in the rotary ring and

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