3M Listeria MDAL96 Manual Del Usuario página 9

5. Verify that the temperature of the 3M Molecular Detection Heat Block Insert is at 100 ±1°C. Place the rack of LS tubes in the 3M Molecular
Detection Heat Block Insert and heat for 15 ±1 minutes
considered a potential biohazard and should NOT be inserted into the 3M Molecular Detection Instrument.
6. Remove the rack of LS tubes from the heating block and allow to cool in the 3M Molecular Detection Chill Block Insert for 10 ±1 minutes
Remove the rack lid during incubation on the 3M Molecular Detection Chill Block Insert.
7. Remove the rack of LS tubes from the 3M Molecular Detection Chill Block Insert/ 3M Molecular Detection Chill Block Tray system. Replace the lid
on the rack of LS tubes and firmly invert 3-5 times to mix. Suspension has to flow freely inside the tube.
8. Firmly tap the lysis tubes rack on the laboratory bench 3-5 times.
9. Place the rack on the laboratory bench. Let it sit undisturbed for 5-10 minutes to allow the resin to settle. Do not mix or disturb the resin at the
bottom of the tube.
99-101ºC
(a) Alternatives to equilibrate the LS tubes to room temperature are to incubate the LS tubes in a 37 ±1°C incubator for 1 hour or at room
temperature overnight (16-18 hours).
(b) An alternative to using dry heat for the lysis step is to use a water bath at 100 ±1°C. Ensure that sufficient water is used to cover up to the
liquid level in the LS tubes. Place the rack of LS tubes in the water bath at 100 ±1°C and heat for 15 ±1 minutes.
(c) The LS solution may freeze when processing less than 48 LS tubes. Freezing of the LS solution will not affect your test. If freezing is observed,
allow the LS tubes to thaw for 5 minutes before mixing.
AMPLIFICATION
1. One Reagent tube is required for each sample and the NC.
1.1 Reagent tube strips can be cut to desired tube number. Select the number of individual Reagent tubes or 8-tube strips needed.
1.2 Place Reagent tubes in an empty rack.
1.3 Avoid disturbing the reagent pellets from the bottom of the tubes.
2. Select 1 Reagent Control (RC) tube and place in rack.
3. To avoid cross-contamination, decap one Reagent tube strip at a time and use a new pipette tip for each transfer step.
4. Transfer lysate to Reagent tubes and RC tube as described below:
Transfer each sample lysate into individual Reagent tubes first followed by the NC. Hydrate the RC tube last.
WARNING: Care must be taken when pipetting LS, as carry-over of the resin may interfere with amplification.
4.1 Use the 3M™ Molecular Detection Cap/Decap Tool-Reagent to decap the Reagent tubes –one Reagent tube strip at a time. Discard cap.
4.2 Transfer 20 µL of Sample lysate from the upper portion of the fluid in the LS tube into corresponding Reagent tube. Dispense at an angle to
avoid disturbing the pellets. Mix by gently pipetting up and down 5 times.
4.3 Repeat step 4.2 until individual Sample lysate has been added to a corresponding Reagent tube in the strip.
4.4 Cover the Reagent tubes with the provided extra caps and use the rounded side of the 3M Molecular Detection Cap/Decap Tool-Reagent to
apply pressure in a back and forth motion ensuring that the cap is tightly applied.
4.5 Repeat steps 4.1 to 4.4 as needed, for the number of samples to be tested.
4.6 When all sample lysates have been transferred, repeat 4.1 to 4.4 to transfer 20 µL of NC lysate into a Reagent tube.
4.7 Transfer 20 µL of NC lysate into a RC tube. Dispense at an angle to avoid disturbing the pellets. Mix by gently pipetting up and down 5 times.
5. Load capped tubes into a clean and decontaminated 3M Molecular Detection Speed Loader Tray. Close and latch the 3M Molecular Detection
Speed Loader Tray lid.
. Samples that have not been properly heat treated during the assay lysis step may be
(b)
0-20ºC 20-25ºC
8
3X
20-25ºC
3X
20-25ºC
20 µL
(English)
.
(c)