hygiena BAX-Q7 Ready Reference for Real Time PCR Assays Installation Guide

hygiena BAX-Q7 Ready Reference for Real Time PCR Assays Installation Guide

STEP 1: PREPARATION

• Add 150 μL protease to 12 mL lysis buffer

• Add 200 µL lysis reagent to cluster tubes

  • Lysis reagent can be stored a 2-8°C for up to two weeks
    

• Ensure thermal blocks are pre-heated to 37°C and 95°C prior to use

• Ensure cooling blocks are stored at 2 – 8°C prior to use
  

STEP 2: LYSIS

• Transfer 5 μL* enriched samples to cluster tubes

  • *For E. coli O157:H7 and STEC, use 20 µL
    

• Heat cluster tubes (First Stage)

  * 37°C for 20 minutes:
  * Campylobacter
  * E. coli O157:H7 Exact
  * E. coli – STEC suite
  * Salmonella
  * Shigella
  * Vibrio

• Heat cluster tubes (Second Stage)
  **95°C for 10 minutes:
  All targets

**

• Cool cluster tubes for a minimum of 5 minutes in cooling block Unopened processed lysates can be stored at 2-8°C for up to two weeks.
  

STEP 3: PCR

• Create rack file, turn on cycler, and initialize
  

• Arrange PCR tubes in PCR cooling block with black carrying tray
  

• Hydrate PCR tablets with 30 µL lysate from cooled lysates
  For Real-Time Salmonella and E. coli O157:H7 Exact, let hydrated tablets sit in the cooling block for 10-30 minutes prior to placing tubes in Q7 Cycler.
  

• On software, click next, place PCR tubes in Q7 cycler and run program
  

• Review results on screen
    Negative
   Positive
   Indeterminate
   Signal error

Ready Reference for Real-Time Listeria PCR Assays

STEP 1: PREPARATION

• Add 150 μL protease to 12 mL lysis buffer
• Add 200 µL Lysing Agent 2 to protease and lysis buffer mixture
• Add 200 µL lysis reagent to cluster tubes
• Ensure thermal blocks are pre-heated to 55°C and 95°C prior to use
• Ensure cooling blocks are stored at 2 – 8°C prior to use

STEP 2: LYSIS

• Transfer 5 μL enriched samples to cluster tubes

• Heat cluster tubes (First Stage)
  55°C for 30 minutes: Genus Listeria L. monocytogenes

• Heat cluster tubes (Second Stage)
  95°C for 10 minutes: All targets

• Cool cluster tubes for a minimum of 5 minutes in cooling block
  Unopened processed lysates can be stored at 2-8°C for up to two weeks

STEP 3: PCR

• Create rack file, turn on cycler, and initialize

• Arrange PCR tubes in PCR cooling block with black carrying tray

• Hydrate PCR tablets with 30 µL lysate from cooled lysates
  Recommended: 10 – 30 min hold in cold block for hydrated tablets prior to placing in Q7 cycler

• On software, click next, place PCR tubes in Q7 cycler and run program

• Review results on screen
  Negative
  Positive
  Indeterminate
  Signal error

For technical support contact us at diagnostics.support@hygiena.com or by visiting www.hygiena.com/technical-support-request

References

• Support | Hygiena

Read User Manual Online (PDF format)

Read User Manual Online (PDF format)  >>

Download This Manual (PDF format)

Download this manual  >>