AOAC 2019 First Action Methods

( 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.

F. PCR Preparation [Before beginning the procedure, don a clean pair of powder-free latex or nitrile gloves.] ( a ) 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. ( b ) Use the following steps to remove the transportation caps from the strips: ( 1 ) Tap on the strips on the bench to ensure the pellets are on the bottom of the tubes.

( 2 ) Carefully open the caps to prevent spilling the freeze-dried pellet.

( 3 ) Visually check that the freeze-dried pellets are present at the bottom of each tube.

( c ) Using a 10 μL Biotix filter pipet tip with a single or multichannel pipet, transfer 10 μL 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 contr ol procedure, use 10 μL of control buffer instead of lysed sample.

( d ) 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. ( e ) Place the GENE-UP® PCR Tube Holder containing the PCR tubes in the plate centrifuge. ( f ) Balance the centrifuge. ( g ) Spin for 10 s.