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PCR, qPCR

About Strategic Alliance
BIOREBA AG, Switzerland, and Qualiplante SAS, France, are pleased to announce they have entered into a global strategic alliance to jointly offer to the market a wide range of products for plant pathogen diagnostic testing based on molecular biology.
 
Benefits of PCR and qPCR tests powered by Qualiplante & BIOREBA:
- Ready-to-use sets
- Optimized for your application
- Validated (sensitivity, specificity, repeatability, reproducibility)
- Lot-to-lot consistent
- Certificate of Analysis (CoA) included
- One-step-reactions
- Positive and negative controls included
- Internal Control IC host gene or IC RNA included
- Long shelf life
- Multiplex with high specificity
For more information please click Product Catalog_Molecular Diagnostic.pdf

PCR methods used for BIOREBA PCR tests,

powered by Qualiplante and by BIOREBA

 

Classical PCR methods

 
End-Point PCR
         
End-Point PCR is the “classical” PCR method,
whereby the DNA is detected after completion
of PCR amplification.
The presence or absence of the corresponding
DNA product is determined by gel
electrophoresis using staining of separated
DNA fragments with a fluorescent dye, and
digital imaging densitometry to visualize DNA bands.                         
     
         
 
End-Point RT-PCR
         
      End-Point RT-PCR is the “classical” PCR method
using RNA as starting material. In a onestep
reaction first, the RNA is reverse transcribed
(RT) into cDNA, then the amplified
cDNA is detected after completion of PCR
amplification.
The presence or absence of the corresponding
DNA product is determined by gel electrophoresis using staining of separated
DNA fragments with a fluorescent dye, and digital imaging densitometry to
visualize DNA bands.
     
         
 
Nested End-Point PCR
         
  Nested End-Point PCR is based on the “classical”
End-Point PCR method. Nested End-Point PCR
reaction involves two distinct sets of primers used
in two successive PCR runs. In the second PCR,
a target within the first PCR product is amplified.
This method allows to reduce non-specific
binding and in parallel to run more total cycles
to increase the amplification.
The presence or absence of the corresponding DNA product is determined
by gel electrophoresis as in a normal End-Point PCR.
     
         
 
Two-Step End Point RT-PCR
         
     The Two-Step End-Point RT-PCR is based
on the End-Point RT-PCR method using RNA
as starting material. The difference is based
on the fact that the overall
reaction is split
into two separate reaction steps (Two-Step).
In a first step the RNA is reverse transcribed
(RT) into cDNA. In a second step, the cDNA is
amplified and analyzed as described above in “End-Point PCR”.                     
     
         
 

Real time PCR (qPCR) methods

 
Taq-Man® qPCR
         
Taq-Man® qPCR is a PCR method, whereby the
amplified DNA is measured in real-time after
each amplification
cycle using fluorescent Taq-
Man® probes.
Taq-Man® probes are oligonucleotides, specific
to the target DNA sequence, that have a
fluorescent probe attached to the 5’ end and a
quencher to the 3’ end. During PCR amplification the fluorophore will be cleaved
away and is decoupled from the quencher. This leads to a strong increase of fluorescence
intensity, which is proportional to the specific target DNA amplification.
The fluorescence is directly measured and quantified in the real-time thermocycler.
Distinct fluorophores can be used to detect multiple targets (Multiplex).
     
         
 
Taq-Man® RT-qPCR
 
         
                   Taq-Man® RT-qPCR uses RNA as starting material.
In a one-step reaction, the RNA is reverse
transcribed (RT) into cDNA, then the cDNA is
amplified during the same reaction and the
fluorescence
of Taq-Man® probes is directly
measured and quantified in the real-time thermocycler.                                                                        
     
         
 
 
SYBR®-Green qPCR
 
         
SYBR®-Green qPCR is a PCR method, whereby
the amplified DNA is measured in real-time
after each amplification cycle using the fluorescent
dye SYBR®-Green.
SYBR®-Green binds only to double-stranded
DNA. When SYBR®-Green binds to the doublestranded
DNA of the PCR products, it emits
fluorescence. This leads to a strong increase in intensity of fluorescence,
which
is proportional to the specific target DNA amplification. Since SYBR®-Green
is not able to bind to single-stranded DNA, the fluorescence
signal fast and
rapidly decreases during the DNA denaturation
step which allows a melting
curve analysis.
     
         
 
 
SYBR®-Green RT-qPCR
 
         
SYBR®-Green RT-qPCR uses RNA as starting
material. In a one-step reaction first, the RNA
is reverse transcribed (RT) into cDNA, then the
cDNA is amplified during the same reaction
and the fluorescence of bound SYBR®-Green
dye is directly measured and quantified in the
real-time thermocycler.                                                                              
     
         
Triplex End-Point PCR - powered by Qualiplante
Art. Nr.: 7AgVitP2
Mengeneinheit: 1 set
Triplex End-Point PCR - powered by Qualiplante
Art. Nr.: 7AgVitP9
Mengeneinheit: 1 set
Nested End-Point PCR - powered by Qualiplante
Art. Nr.: 7AlmWBP2
Mengeneinheit: 1 set
Nested End-Point PCR - powered by Qualiplante
Art. Nr.: 7AlmWBP9
Mengeneinheit: 1 set
Nested End-Point PCR - powered by Qualiplante
Art. Nr.: 7AP---P2
Mengeneinheit: 1 set
Nested End-Point PCR - powered by Qualiplante
Art. Nr.: 7AP---P9
Mengeneinheit: 1 set
One-Step RT-PCR - powered by Qualiplante
Art. Nr.: 7ApMV-P2
Mengeneinheit: 1 set
One-Step RT-PCR - powered by Qualiplante
Art. Nr.: 7ApMV-P9
Mengeneinheit: 1 set
Multiplex End-Point PCR (without RT) with IC - powered by Qualiplante
Art. Nr.: 77VV--P9w
Mengeneinheit: 1 set