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About PCR

BioChek provides diagnistics PCR tests for swine and poultry, as well as a range of PCR accessories such as standards.

Read more about our PCR test kits below.

PCR:

  • With PCR you are making copies of a specific DNA sequence
  • With enough copies you can perform measurements
  • PCR is very sensitive
  • Proper laboratory technique is important
  • Running the PCR on the thermocycler
  • Be aware of contamination risks

BioChek PCR in a nutshell:

  • Creating a work list/plate lay-out
  • Preparation of reaction mix
  • DNA/RNA extraction
  • Adding DNA/RNA to the reaction mix
  • Running the PCR on the thermocycler
  • Analysing the results, creating reports

Reagents needed to perform a PCR reaction

  • Nucleotides, included in the mastermix
  • DNA polymerase (taq), included in the mastermix
  • Buffer, included in the mastermix
  • Magnesium, included in the mastermix
  • Primers, included in the primer/probe/IC mix
  • Probes/beacons, included in the primer/probe/IC mix
  • Internal control, included in the primer/probe/IC mix

PCR or ELISA?

  • PCR is capable of early detection
  •  Pathogen quantification possible with PCR
  •  PCR is less likely to cross react
  •  PCR detects the pathogen directly (ag instead of ab)
  • Using both ELISA and PCR alternately can be benificial

BioChek PCR kit design

  • Primers and probe are specific for a unique sequence
  • Targeted DNA sequence determines primer/probe properties
    • All BioChek PCR kits can be used simultaneously
    • DNA/RNA targets can also be combined
    • All targets can be quantified using standards

PCR trouble shooting

  • Inhibited samples:
    • Repeat sample diluted (5-10 x). Be aware: sensitivity will be decreased
    • Extract DNA using a different method
  • Possible contamination , negative controls positive:
    • Clean work surfaces with a nucleic acid degrading solution
    • Clean equipment with a nucleic acid degrading solution
    • Repeat test
  • Still positive NC controls:
    • Perform wipe tests of work surfaces and equipment
    • Wipe tests will detect DNA traces and amplicons
  • Don’t report results while you suspect a contamination, resume reporting after you have found and solved the problem