R 3

02 0

4 2

1 (8

)

Pre-incubation: 1 cycleStep 1: 40 °C for 2 minStep 2: 95 °C for 10 min

Amplifi cation: 45 cyclesStep 1 : 95 °C for 15 secStep 2*: 63 °C for 30 sec

* Fluorescence detection1 cycle

2 min 10 min 15 sec | 30 sec*|

45 cycles

95 °C

63 °C

40 °C

Brucella Detection Kit Quick Reference Procedure

PCR kit for the qualitative detection of Brucella abortus, Brucella melitensis and Brucella spp. DNA using real-time PCR instruments.

Before starting, it is strongly recommended to read the entire product manual available on our website.

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

u FAM (Brucella abortus), HEX (Brucella melitensis), ROX (Brucella spp.) and Cy5 (Internal Control).

For some real-time PCR instruments the probe quencher as well as the usage of a passive reference dye has to be specified. This kit contains probes with DABCYL as quencher and no passive reference dye.A Color Compensation is necessary for users of the LightCycler® 480 System: Color Compensation Set 3 (Order No. A 500 10).

Order no. R 302 04

- 5‘Nuclease -

Version 5, November 2017

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

foodproof®

FAM HEX ROX Cy5 Result Interpretation

+ + + + or - Positive for Brucella abortus, Brucella melitensis and Brucella spp.

- + + + or - Positive for Brucella melitensis and Brucella spp.

+ - + + or - Positive for Brucella abortus and Brucella spp.

- - + + or - Positive for Brucella spp.

- - - + Negative for Brucella

- - - - Invalid

foodproof® is a registered trademark of BIOTECON Diagnostics. The above mentioned real-time PCR instruments are registered trademarks of their respective holders.

foodproof® Brucella Detection Kit

R 302 04Kit for 96 reactionsStore kit at -15 to -25 °C

For food testing purposesFOR IN VITRO USE ONLYMade in Germany

BIOTECON Diagnostics GmbH | Hermannswerder 17, 14473 Potsdam, Germany | +49 331 2300 200 | www.bc-diagnostics.com

3. ADD SAMPLES AND CONTROLSPipet 5 µl of samples, negative control (colorless cap) or

Control Template (purple cap) into respective wells.

2. ADD PCR MIXPipet 20 µl of prepared PCR mix into each strip or plate well.

1. PREPARE PCR MIXAdd 18 µl of Master Mix (yellow cap), 1 µl Enzyme Solution (red cap) and 1 µl Internal Control (white cap) 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.

PREPARATION OF THE PCR MIXTake appropriate precautions to prevent contamination, e.g. by using fi lter tips and wearing gloves. Thaw reagents, mix (do not vortex!), and briefl y spin vials before opening.

4. SEALSeal strips/plate accurately.

5. CENTRIFUGEBriefl y spin strips/plate in a suitable centrifuge.

6. START REAL-TIME PCR RUNCycle samples as described above.

+20 µl

+5 µl

+18 µl + 1 µl + 1 µl