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5.
Verify that the temperature of the 3M Molecular Detection Heat Block Insert is at 100 ±1°C. Place
1
the rack of LS tubes in the 3M Molecular Detection Heat Block Insert and heat for 15 ±1 minutes
2
(b)
. Samples that have not been properly heat treated during the assay lysis step may be considered a
3
potential biohazard and should NOT be inserted into the 3M Molecular Detection Instrument.
4
6.
Remove the rack of LS tubes from the heating block and allow to cool in the 3M Molecular
5
Detection Chill Block Insert for 10 ±1 minutes
(c)
.
Remove the rack lid during incubation on the
6
3M Molecular Detection Chill Block Insert
.
7
7.
Remove the rack of LS tubes from the 3M Molecular Detection Chill Block Insert/ 3M Molecular
8
Detection Chill Block Tray system. Replace the lid on the rack of LS tubes and firmly invert 3-5
9
times to mix.
Suspension has to flow freely inside the tube.
10
8.
Firmly tap the lysis tubes rack on the laboratory bench 3-5 times.
11
9.
Place the rack on the laboratory bench. Let it sit undisturbed for at least 5 minutes to allow the resin
12
to settle.
Do not mix or disturb the resin at the bottom of the tube
.
13
14
15
16
(a)
Alternatives to equilibrate the LS tubes to room temperature are to incubate the LS tubes in a 37
17
±1°C incubator for 1 hour or at room temperature overnight (16-18 hours).
18
(b)
An alternative to using dry heat for the lysis step is to use a water bath at 100 ±1°C. Ensure that
19
sufficient water is used to cover up to the liquid level in the LS tubes. Place the rack of LS tubes
20
in the water bath at 100 ±1°C and heat for 15
±1
minutes.
21
(c)
The LS solution may freeze when processing less than 48 LS tubes. Freezing of the LS solution
22
will not affect your test. If freezing is observed, allow the LS tubes to thaw for 5 minutes before
23
mixing.
24
25
J. A
MPLIFICATION
26
1.
One Reagent tube is required for each sample and the NC.
27
1.1
Reagent tubes strips can be cut to desired tube number. Select the number of individual Reagent
28
tubes or 8-tube strips needed.
29
1.2
Place Reagent tubes in an empty rack.
30
1.3
Avoid disturbing the reagent pellets from the bottom of the tubes.
31
2.
Select 1 Reagent Control (RC) tube and place in rack.
32
3.
To avoid cross-contamination, decap one Reagent tubes strip at a time and use a new pipette tip for
33
each transfer step.
34
4.
Transfer lysate to Reagent tubes and RC tube as described below:
35
36
Transfer each sample lysate into individual Reagent tubes
first
followed by the NC. Hydrate the RC
37
tube
last
.
38
WARNING:
Care must be taken when pipetting LS, as carry-over of the resin may interfere with
39
amplification.
40
4.1
Use the 3M™ Molecular Detection Cap/Decap Tool-Reagent to decap the Reagent tubes –one
41
Reagent tubes strip at a time. Discard cap.
42
4.2
Transfer 20 µL of Sample lysate from the upper portion of the fluid in the LS tube into
43
corresponding Reagent tube. Dispense at an angle to avoid disturbing the pellets. Mix by gently
44
pipetting up and down 5 times.
45
Candidates for 2016 Method of the Year
120