PCM124202EN00 (12/20) Direct Swab Preparation – Deepwell Format Amplification and Detection Procedure Open Master Mix slowly. Add 135 μL of Rehydration Solution. Replace cap and allow it to sit for 1-2 minutes. Gently pipette rehydrated Master Mix up and down 3-5 times. Avoid creating bubbles. 12x Master Mix Rehydration Solution Negative Control Positive Control Kit Contents 9 6 Emergency Use Only Process Buffer 1 Pipette 400 μL of the Process Buffer into each deepwell as needed. 4 Heat the deepwell plate for 10 minutes at 95°C at 300 rpm. Amplification and Detection Sample Preparation 5 10 15 20 25 30 35 40 45 mL External Controls To process the samples, add patient swab(s) into each deepwell. Vigorously twirl swab for 10 seconds in Process Buffer. Roll the swab head against the inside of the tube wall as you remove it. Dispose of used swab in biohazard waste container. 3 H G F E D C B A 5 Remove the deepwell plate from the heating block and place onto the frozen cooling block at room temperature for 10 minutes. Direct Swab with Deepwell Format Procedure Card Insert PCR plate into appropriately programmed thermocycler and select appropriate Lyra assay protocol. Approximately 70 minutes per run. 5 10 15 20 25 30 35 40 45 mL H G F E D C B A 2 To process the controls add 50 μL of Positive and Negative Control to the designated deepwell. H G F E D C B A H G F E D C B A Mix each deepwell (column or row) 3 times using a pipette set at 150uL. Immediately transfer 5 μL of sample to the PCR plate. Repeat as needed. Seal the PCR plate. Centrifuge PCR plate for 15 seconds. 8 H G F E D C B A Study the Package Insert and Technical Bulletin thoroughly before using this Procedure Card. This is not a complete Package Insert. A B C D E F G H Place the microwell PCR plate on the frozen microwell plate cooler. Add 15 μL of Master Mix to each well. 7