foodproof^® SL Clostridium perfringens Detection Kit

PROGRAM SETUP

Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:

• FAM (α-toxin gene) and VIC/HEX (Internal Control).

PCR Cycling Conditions:

• Pre-incubation: 1 cycle, Step 1: 95 °C for 10 min
• Amplification: 40 cycles, Step 1: 95 °C for 15 s, Step 2*: 60 °C for 60 s

* Fluorescence detection

DATA INTERPRETATION

Verify results of positive (Control Template) and negative (H₂O) controls, before interpreting the sample results. Always compare samples to positive and negative controls. Review data from each channel and interpret results as described in the table.

PREPARATION OF THE RT-PCR MIX

Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves. Thaw reagents, mix (do not vortex!) and briefly spin vials before opening.

1. PREPARE PCR MIX

   Add 10 µL Master Mix (2X M) and 4 µL Primer/Probe Mix (P) for each reaction to a suitable tube. (n samples + 2 controls + at least one additional reaction to cover pipetting loss). Mix carefully but thoroughly by pipetting up and down.

2. ADD PCR MIX

   Pipet 14 µL of prepared PCR mix into each strip or plate well.

3. ADD SAMPLES AND CONTROLS

   Pipet 6 µL of samples, negative control (PCR-grade H₂O, not included) or Control Template (C) into respective wells.

4. SEAL

   Seal strips/plate accurately. [Seal icon]

5. CENTRIFUGE

   Briefly spin strips/plate in a suitable centrifuge. [Centrifuge icon]

6. START REAL-TIME PCR RUN

   Cycle samples as described above. [Play icon]