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    R 302 80 21 microproof-SARS-CoV-2-Screening

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    R_302_80_21_microproof-SARS-CoV-2-Screening

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    R 302 80 21 microproof-SARS-CoV-2-Screening

    Uploaded by

    smzaheer_alam
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    R_302_80_21_microproof-SARS-CoV-2-Screening

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    R_302_80_21_microproof-SARS-CoV-2-Screening

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    R_302_80_21_microproof-SARS-CoV-2-Screening

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    2 views2 pages

    R 302 80 21 microproof-SARS-CoV-2-Screening

    Uploaded by

    smzaheer_alam
    R_302_80_21_microproof-SARS-CoV-2-Screening

    Copyright:

    © All Rights Reserved
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    of 2

    microproof®

    SARS-CoV-2 Screening Kit


    Quick Guide
    - 5‘Nuclease -
    Order no. R 302 80 (L) Version 1, May 2020

    PCR kit for the qualitative detection of the E gene (SARS-related coronavirus) using real-time PCR instruments.
    Before starting, it is strongly recommended to read the entire product manual available on our website.
    The Process Control (white cap) contains a stabilized solution of bacteriophage MS2 to be used as
    preparation/internal amplification control. Before starting the RNA extraction procedure, it is recommended
    to add 5 µl Process Control to each sample or the respective volume to the prepared lysis mix of the Magnetic
    Preparation Kit VI (order no. S 400 20.1 L), the Sample Preparation Kit IV (order no. S 400 16) or other suitable
    extraction kits.

    PROGRAM SETUP
    Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:
    u FAM (E gene; SARS-related coronavirus) and ROX (Process Control).

    95 °C Reverse Transcription: 1 cycle


    Step 1: 50 °C for 30 min
    Pre-incubation: 1 cycle
    60 °C
    Step 2: 95 °C for 5 min
    50 °C Amplification: 45 cycles
    30 min | 5 min 5 sec | 40 sec* Step 1 : 95 °C for 5 sec
    Step 2*: 60 °C for 40 sec
    1 cycle 1 cycle 45 cycles
    * Fluorescence detection
    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 a non-fluorescent “dark” quencher and no passive reference dye.

    DATA INTERPRETATION
    Verify results of positive (Control Template) and negative (Negative Control) controls, 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.

    FAM ROX Result Interpretation

    + + or -
    R 302 80 21 (2)

    Positive for SARS-related coronavirus


    - + Negative for SARS-related coronavirus

    - - Invalid (Note: Expected result if H2O PCR-grade is used as template)

    microproof® is a registered trademark of BIOTECON Diagnostics.


    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.
    Reverse transcriptase is a very temperature-sensitive enzyme. It is recommended to use a cooling block (at
    4 °C) to avoid degradation of the Enzyme Solution (red cap).
    After usage, store the enzyme solution immediately at -15 to -25 °C.

    1. PREPARE RT-PCR MIX


    Add 14 µl of Master Mix (yellow cap) and 1 µl Enzyme Solution (red 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. +14 µl + 1 µl

    2. ADD RT-PCR MIX


    Pipet 15 µl of prepared RT-PCR mix into each strip or plate well.
    +15 µl

    3. ADD SAMPLES AND CONTROLS


    Pipet 10 µl of samples, Negative Control (orange cap) or
    Control Template (purple cap) into respective wells.
    +10 µl

    4. SEAL
    Seal strips/plate accurately.

    5. CENTRIFUGE
    Briefly spin strips/plate in a suitable centrifuge.

    6. START REAL-TIME PCR RUN


    Cycle samples as described above.

    microproof® R 302 80 (L) Made in Germany


    SARS-CoV-2 Kit for 96 (480) reactions
    Screening Kit Store kit at -15 to -25 °C
    BIOTECON Diagnostics GmbH | Hermannswerder 17, 14473 Potsdam, Germany | +49 331 2300 200 | www.bc-diagnostics.com

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