Mycoplasma pneumoniae Chlamydophila pneumoniae - Mikrogen

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DIAGENODE MYCOPLASMA PNEUMONIAE / CHLAMYDOPHILA PNEUMONIAE REAL-TIME PCR USER MANUAL
III . PCR Reaction Set-up
(ABI 7000-7300-7500-7900/ Roche Lc480-Lc2.0 / Biorad iCycler-IQ5-CFX96 / Stratagene MX3000P-
3500P / Qiagen Rotor-Gene / Cepheid SmartCycler II (on request) )
With Quantifast Multiplex + R kit (ref : 204756), Qiagen (Not supplied)
NB:
For the Qiagen Rotor-Gene, we recommend the Rotor-Gene Multiplex PCR Kit, Qiagen (Not
supplied) (Ref: 204774).
For the LightCycler 2.0 (Lc2.0), we specially suggested LightCycler ® TaqMan ® Master, Roche (Not
supplied) (ref: 04535286001).
• PCR 50 μl:
(ABI 7000-7300-7500-7900/ Roche Lc480 / Biorad iCycler-IQ5 / Stratagene MX3000P-
3500P / Qiagen Rotor-Gene)
For the Qiagen Rotor-Gene, we recommend the Rotor-Gene Multiplex PCR Kit, Qiagen
(Not supplied) (Ref: 204774).
25 μl ................................... 2X master mix
5 μl ..................................... Diagenode Mycoplasma pneumoniae and Chlamydophila pneumoniae
double-dye probes & primers
5 μl ..................................... Diagenode DNA extraction & PCR inhibition control double-dye probe &
primers
10 μl ................................... Human sample (or Mycoplasma pneumoniae and Chlamydophila
pneumoniae positive control)
5 μl ..................................... Diagenode water
50 μl ................................... Final volume
NB:
With ABI system, you can add 1 μl of the ROX solution (Qiagen)
• PCR 25 μl:
(ABI 7000-7300-7500-7900/ Roche Lc480-Lc2.0 / Biorad iCycler-IQ5-CFX 96 / Stratagene
MX3000P-3005P / Qiagen Rotor-Gene / Cepheid SmartCycler II (on request) )
12.5 μl ................................ 2X master mix
2.5 μl .................................. Diagenode Mycoplasma pneumoniae and Chlamydophila pneumoniae
double-dye probes & primers
2.5 μl .................................. Diagenode DNA extraction & PCR inhibition control double-dye probe &
primers
5 μl ..................................... Human sample (or Mycoplasma pneumoniae and Chlamydophila
pneumoniae positive control)
2.5 μl .................................. Diagenode water
25 μl ................................... Final volume
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