Microbiology Methods for ERP Review 3-2020

65

GENE ‐ UP ® SLM 2 Collaborative Study

July 2019

c) Clip the GENE - UP® Lysis Tube Holder on the GENE - UP® Heavy Rack Holder. d) Transfer 20 μ L of sample into the lysis tube. Use the Plate Map from the GENE - UP® Routine software to pipet each sample into the correct plate position. e) Remove the GENE - UP® Lysis Tube Holder from the GENE - UP® Heavy Rack Holder. f) Clip the GENE - UP® Lysis Tube Holder on the Troemner Vortex Mixer Adaptor. g) Run the vortex mixer at 2200 rpm for 5 min. Note: When using the Vortex - Genie® Pulse, fit it with the GENE - UP® Lysis Rack Adaptor. Run the vortex at maximum speed for 5 min. The maximum speed must be above 2000 rpm. h) When lysis is complete, remove the GENE - UP® Lysis Tube Holder from the Troemner Vortex Mixer Adaptor. i) Clip the GENE - UP® Lysis Tube Holder into the GENE - UP® Heavy Rack Holder and proceed to Final Setup for PCR. Note: The lysate can be stored for up to 3 days at 2–8°C or at  ‐ 15 to  ‐ 31°C for extended storage. j) PCR Preparation a) Before beginning the procedure, don a clean pair of powder ‐ free latex or nitrile gloves. b) Use the plate map created in the GENE - UP® Routine software to determine the number of PCR tubes required from the GENE - UP® PCR Kit and place the correct number of PCR tubes in the GENE - UP® PCR Tube Holder. If less than eight tubes in a strip are required, the strips can be cut apart, and only the used tubes are placed in the GENE - UP® PCR Tube Holder. Note: Only remove the required number of strips from the pouch and carefully reseal the pouch after opening. c) Use the following steps to remove the transportation caps from the strips: d) Tap on the strips on the bench to ensure the pellets are on the bottom of the tubes. e) Carefully open the caps to prevent spilling the freeze - dried pellet. f) Visually check that the freeze - dried pellets are present at the bottom of each tube. g) Using a 10 μ L Biotix filter pipette tip with a single or multichannel pipette, transfer 10 μ L of lysed sample (red) in the appropriate PCR tube. To determine the appropriate plate position for each sample, refer to the Plate Map from the GENE - UP® Routine software. Note: Do NOT agitate the lysate before aspirating the sample. The solid material must stay at the bottom of the tube. Note: Visually check the tips to confirm the absence of beads, bubbles, and for correct volume of lysate. Note: For negative control procedure, use 10 μ L of control buffer instead of lysed sample. h) Place and seal the strip caps onto each strip tube using the GENE - UP® Lysis Tube Remover Tool. If less than eight caps are required, the caps can be cut apart, and only the used caps are placed onto the strip tubes.

10