Mycobacterium Tuberculosis - Diagenode Diagnostics

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DIAGENODE MYCOBACTERIUM TUBERCULOSIS REAL-TIME PCR USER MANUAL
Troubleshooting guide
> Nucleic Acid isolation
Refer to the manufacturer’s product insert
NB:
Perform preventive maintenance of workstations for automated extraction (according to the
manufacturer’s recommendations).
Check the expiration date and storage conditions for solutions provided
> PCR
The signal detected for the positive Mycobacterium tuberculosis control, the DNA extraction
& inhibition control (DIA-EIC/DNA-050) are negative or weak.
The PCR conditions do not comply with the protocol:
1. Check extraction procedure.
It’s important to select an appropriate extraction procedure. The extraction procedure
must be validated and limit the presence of inhibitory factors.
2. Check PCR reaction set-up.
To obtain a high sensitivity, you must respect volumes described and the master mixes
suggested.
NB:
Check the calibration of pipettes used (according to the manufacturer’s recommendations).
Check the expiration date and storage conditions of master mixes suggested.
3. Check the PCR profile.
To obtain a high sensitivity, respect the protocol adapted to the real-time PCR used.
4. Check dyes selected.
Check and select dyes adapted for each real-time PCR platform.
5. Interpretation of results.
Check the table and select the appropriate channel.
Important note:
Check storage conditions (during and after experiments) and expiration date.These two
conditions highly influence results obtained.
> Real-time PCR system
Refer to the manufacturer’s product insert.
NB:
Perform maintenance of the real-time PCR platform (according to the manufacturer’s
recommendations).
For trouble shootings/questions, please contact us:
Tel.: +32 (0)4 364 20 50 - info@diagenodediagnostics.com
Europe Diagenode sa / CHU - Tour GIGA - B34 - 3 e étage // Avenue de l’Hôpital, 1 // 4000 Liège (Sart Tilman) // Belgium // Phone: (+32) 4 364 20 50 // Mail: info@diagenode.com