Device Settings - R-Biopharm RIDAGENE CMV PG9005 Bedienungsanleitung

9.2 Preparation of the PCR mix for quantitative detection
The total number of the reactions needed for PCR (samples and control reactions)
must be calculated. In addition to the standards, one positive and one negative
control must be included in each test run.
Pipette 20 µl of the master mix into each reaction vial (vial/plate).
Negative control:
Sample:
Positive control:
Standard (A, B, C, D):
Seal the reaction vials or plates, briefly centrifuge at slow speed, and transfer into the
real-time PCR device. Start PCR according to device settings (see Table 3).

9.3 Device settings

Table 3: Real-time PCR profile for LightCycler
CFX96
Decontamination
Initial denaturation
Cycles
PCR Denaturation
Annealing
Extension
Note: For the standards A, B, C, and D, enter the total number of copies for
each reaction into the setup file of the software program of the real-time
PCR device. An amount of 20 µl standard is used, resulting in the
following concentrations:
Standard A: 1 x 10² gEq/reaction
Standard B: 1 x 10
Standard C: 1 x 10
Standard D: 1 x 10

RIDA GENE CMV
Add 20 µl PCR water to the pre-pipetted master mix as a
negative control.
Add 20 µl DNA extract to the pre-pipetted master mix of
the sample reactions.
Add 20 µl Positive Control for the pre-pipetted master
mix to the reaction vial.
Add 20 µl Standard (A, B, C, D) for each pre-pipetted
master mix to the designated reaction vials.
3
gEq/reaction
4
gEq/reaction
5
gEq/reaction
®
480II, Mx3005P, Rotor-Gene Q, and
2 min, 50 °C
2 min, 94 °C
45 cycles
10 sec, 94 °C
30 sec, 60 °C, "detection"
20 sec, 72 °C
2019-05-28
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