Realtime PCR

Highly sensitive and fast method using specific primers for the detection of RNA-or DNA containing pathogens. The detection is followed by fluorescence measurements in real time during the amplification reaction with a Realtime PCR Cycler.

When to use Realtime PCR

  • Screening of non-symptomatic plants
  • Determination of infection levels of a certain population
  • Screening of samples with known lower pathogen titer (e.g. seeds)
  • Diagnostic method with good confidence in negative samples

Advantages and Limitation

      • Fast: 30 – 60 min
      • Quantitation possible
      • Highly specific
      • Very sensitive
      • Expensive laboratory equipment needed
We are now offering our complete LOEWE®REALTIME reaction kits for the detection plant pathogens. The kits utilize the Realtime Polymerase Chain Reaction technique, including pathogen-specific primers in combination with DNA-binding dyes or fluorescently labeled probes to detect the amplification. Depending on the pathogen, we use detection with SolisGreen®, which is compatible with all common qPCR cyclers with standard settings for SYBR®Green. For others FAM™-labeled probes are utilized. Please see details below.                                      * provided by SOLIS Biodyne

LOEWE®REALTIME reaction kits for Bacteria

The reaction (dye-based or  probe-based, depending on the specific kit) is carried out either with SolisGreen® or with FAM-labeled probes for fluorescence detecting. A detailed manual is also included in each kit.
An innovative Internal Control DNA/RNA Assay (LOEWE® Cat. No. 0600IC/100) is additionally available for probe-based LOEWE®REALTIME reaction kits for bacteria. It can be used to verify successful DNA extraction and amplification in all samples in a separate reaction.
Kit Content
  • Primer Mix or Primer/Probe Mix (pathogen specific)
  • 5 x Mastermix*
  • Positive control (Synthesized DNA)
  • Negative control (DNA from healthy plants)
  • PCR-grade water
* provided by SOLIS Biodyne

LOEWE®REALTIME reaction kits for RNA Viruses

(ONE-Tube ONE-Step)

The reaction is carried out in one tube starting with the reverse transcription of plant virus RNA prior to cDNA amplification. Each kit includes specific primers and FAM-labelled probe for reliable and highly sensitive detection of the target pathogen.
Additionally, an internal control is already included, which is based on universal primers specific for plant RNA combined with a HEX-labelled probe. This allows to verify successful RNA extraction and amplification in all samples.
Kit Content
  • Primer/Probe Mix (pathogen specific)
  • 5x One-step Multiplex Probe Mix (ROX)*
  • 40x One-step SOLIScript® Mix*
  • Positive control (Synthesized DNA)
  • Negative control (DNA from healthy plants)
  • PCR-grade water
* provided by SOLIS Biodyne

 

 

LOEWE® Biochemica GmbH

Mühlweg 2a
82054 Sauerlach
Germany
Tel. +49 8104 61620
Fax. +49 8104 61648

 

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