Bio-Rad VINEO™ Brettanomytest PCR Kit - Manual (Page 8)

2.2 After preparation, the reagent mix (amplification and probe solutions)
must be used immediately, or can be stored for 2 hours between +2°C
and +8°C.
2.3 Distribute 45 μl of this reagent mixture per well according to the defined
plate surface. Appendices B and C propose plate surfaces to be used
for the Chromo4™/CFX96™ and MiniOpticon™ thermocyclers
respectively.
Add 5 μl of sample or negative control or Qs PCR positive control and
seal the wells on the plate or the strips hermetically. It is important to
insist on sealing the wells and the strips to avoid any evaporation
phenomenon during the PCR reaction.
It is important to avoid the presence of bubbles at the bottom of the wells
by pipetting cautiously. To eliminate bubbles after sealing the plate or
closing the PCR strips, you may centrifuge the plate of PCR strips briefly.
The plate can be store at +2°C to +8°C for 2 hours.
2.4 Put the plate or the PCR strips into the thermocycler. Make sure that they
are correctly oriented (well A1 at the top left).
Close the reaction module.
3. Starting the amplification reaction
To restart the PCR, refer to the user guide for the Bio-Rad thermocycler for
the VINEO™ Brettanomytest PCR Kit.
10 - ANALYSIS OF THE DATA
Data analysis can be carried out directly at the end of the amplification
reaction or later by re-opening the data file. To open the data files and
analyse the results of the PCR, refer to the user guide for the Bio-Rad
thermocycler for the VINEO™ Brettanomytest PCR Kit.
11 - INTERPRETATION OF THE RESULTS
To obtain the analysis results, you only have to read the values of Ct (Cycle
threshold): value of the amplification cycle from which fluorescence increases
significantly above background noise.
Manual analysis will only be used for a qualitative analysis (presence or
absence).
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