3M™ Molecular Detection System
E. coli O157 (including H7) Protocol Reference Guide
Enrichment
25g
225mL 3M BPW ISO
8–18 hr at 41.5°C (± 1°)
Raw Meat
325g
975mL 3M BPW ISO
10–18 hr at 41.5°C (± 1°)
Food
375g
1500mL 3M BPW ISO
10–18 hr at 41.5°C (± 1°)
Leafy Produce
• 200g in Butterfield’s phosphate buffer (w/w)
• 125g of product rinsate
25g
225mL 3M BPW ISO
18–24 hr at 41.5°C (± 1°)
2X strength 125mL 3M BPW ISO
18–24 hr at 41.5°C (± 1°)
Lysis
LS
1. Transfer 20µL of Sample into individual Lysis (LS) tube.
20µL NC
3X
2. T ransfer 20µL of Negative Control (NC) into one LS tube.
3. C
over rack of LS tubes with lid and invert 3 times to mix.
4. Place rack with LS tubes into a heating block (100°C ± 1°C) and heat
for 15 minutes.
5. R
emove the rack of LS tubes from heating block and remove lid.
Place rack on Chill Block for 10 minutes.
1. T ransfer 20µL of Sample
into individual LS tube.
2. Transfer 20µL of NC
into one LS tube.
99-101ºC
6. R
emove the rack from Chill Block and mix 3 times. Tap rack on
bench 3 times.
7. Let set on bench top at room temperature for 5 minutes.
3. Cover rack with lid.
Invert 3 times to mix.
0-20ºC
4. Heat LS tube rack at
100°C (±1°) for 15 minutes.
Amplification
5. Cool LS tube rack without
lid in Chill Block for 10 minutes.
3X
8. T ransfer 20µL of each sample lysate into Reagent tubes. Mix by gently
pipetting up and down 5 times. Seal tubes with caps.
9. If needed, transfer 20µL of sample lysate into MC tubes. Mix by gently
pipetting up and down 5 times. Seal tubes with caps.
10. Transfer 20µL NC lysate into one Reagent tube. Mix by gently pipetting up
and down 5 times. Seal tube with cap.
3X
6. Cover rack. Invert 3 times to mix.
Firmly tap rack on lab bench 3 times.
20-25ºC
7. Let set on bench top at room
temperature for 5 minutes.
20µL
11. Transfer 20µL NC lysate into one Reagent Control (RC) tube. Mix by gently
pipetting up and down 5 times. Seal tube with cap.
12. Transfer closed tubes to Speed Loader Tray.
8–11. Reagent hydration. Pipette
up and down 5 times to mix.
12. T ransfer closed tubes
to Speed Loader Tray.
13. Place Speed Loader Tray into
Instrument and close the lid.
Results in 75 minutes.
13. P lace Speed Loader Tray into instrument and close the lid to start the assay.
Food Safety
3M Canada
3M Center
Post Office Box 5757
Building 275-5W-05
London, Ontario N6A 4T1
St. Paul, MN 55144-1000
Canada
USA
1-800-364-3577
1-800-328-6553
www.3M.com/3MMolecularDetectionSystem
Please recycle. Printed in U.S.A.
© 3M 2012. All rights reserved.
70-2009-9511-9 Rev-0812
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Used under license in Canada.