plant pathogen booklet
TRANSCRIPT
PLANT PATHOGEN DETECTION
Aspergillus specie fungal conidiospores Page 4
www.norgenbiotek.com
ISO
9001:2008
ISO
13485:2003
ISO
15189:2012
Ordering Information
Website: www.norgenbiotek.com
Phone: 905-227-8848
Toll Free in North America: 1-866-667-4362
Fax: 905-227-1061
Email: [email protected]
Address:
Norgen Biotek Corp.
3430 Schmon Parkway
Thorold, ON
L2V 4Y6
CANADA
PLANT PATHOGEN DETECTION
Principle of the Test
Norgen’s PCR Detection Kits constituent a ready-to-use system for the isolation and detection
of Plant pathogens using end-point or real-time PCR. The kits first allows for the isolation of RNA
or DNA from samples using spin-column chromatography based on Norgen’s proprietary
resin. The RNA or DNA is isolated free from inhibitors, and can then be used as the template in
a PCR or one step RT-PCR reaction for pathogen detection using the provided Detection
Mastermix. The Detection Mastermix contains reagents and enzymes for the specific
amplification of a region of the target genome or RNA.
In addition, Norgen’s PCR Detection Kits contain a second Mastermix, the RT-PCR or PCR
Control Master Mix, which can be used to identify possible PCR inhibition and/or inadequate
isolation via a separate RT-PCR reaction with the use of the provided PCR control (PCRC) or
Isolation Control (IsoC), respectively. The kits are designed to allow for the testing of 24
samples.
Key Features
Rapid isolation of high quality RNA or DNA from challenging plant tissues or culture.
High sensitivity and specificity
Primer set and controls also available separately
Ideal for use in:
1. Identification of plant pathogen (Research only)
2. Epidemiologial Studies
3. Field Surveillance of Pathogens
4. Crop, vegetable and fruit quality assessment
End-Point PCR Real-Time PCR (SYBR® Green) Real-Time PCR (TaqMan®)
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Product Description Cat. # Page
Aspergillus niger
PCR Detection Kit
Isolation and detection of Aspergillus niger DNA
from plant tissue and fruits. 32900 4
Botrytis cinerea
PCR Detection Kit
Isolation and detection of Botrytis cinerea DNA
from plant tissue and fruits. 29400 5
Cladosporium cladosporioides
PCR Detection Kit
Isolation and detection of Cladosporium
cladosporioides DNA from plant tissue and fruits. 33000 6
Penicillium sp.
PCR Detection Kit
Isolation and detection of Penicillium sp. DNA
from plant tissue and fruits. 33200 7
Saccharomyces cerevisiae
PCR Detection Kit
Isolation and detection of Saccharomyces
cerevisiae DNA from plant tissue, fruits and
beverage samples including wine and juice.
33300 8
Plum Pox Virus
RT-PCR Detection Kit
Isolation and detection of Plum Pox Virus RNA
from plant tissue, fruits and sap. 33400 9
Phomopsis viticola
CR Detection Kit
Isolation and detection of Phomopsis
viticola from plant tissues and fruits. 34900 10
Erwinia amylovora
PCR Detection Kit
Isolation and detection of Erwinia
amylovora from plant tissue. 35100 11
TABLE OF CONTENTS
Figure 1. Detection of A. niger using the A. niger PCR Detection
Kit. A representative 1X TAE 1.5% agarose gel showing the
amplification of A. niger positive (lane 1 and 2) negative (lane 3
and 4) controls. The size of the A. niger target amplicon
corresponds to 363 bp as represented by the provided DNA
Marker (M).
A ready-to-use system for the isolation and
detection of A. niger using end-point PCR
Norgen’s Aspergillus niger PCR Detection Kit is a ready-to-
use system for the isolation and detection of A. niger from
plant samples. First, the kit contains components for the
rapid isolation of total DNA, including fungal DNA, from
plant samples using spin-column chromatography based
on Norgen’s proprietary resin. Second, the kit contains A.
niger Master Mix and controls to allow for PCR
amplification. The amplified PCR products are then
detected using agarose gel electrophoresis. Alternatively,
detection can be performed based on real-time PCR
using melt curves.
The A. niger Mastermix contains reagents and enzymes for
the specific amplification of a 363 bp region of the fungal
genome. In addition, Norgen’s A. niger PCR Detection Kit
contains a second Mastermix, the PCR Control Master
Mix, which can be used to identify possible PCR inhibition
and/or inadequate isolation via a separate PCR reaction
with the use of the provided PCR control (PCRC) or
Isolation Control (IsoC), respectively. This kit is designed to
allow for the testing of 24 samples. The Aspergillus
niger PCR Primer Set and Controls are also available
separately for end-point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range of Norgen’s Aspergillus niger PCR
Detection Kit was determined by analyzing a dilution
series of an A. niger quantification standards ranging
from 1 x 106 cfu/µL to 1 x 101 cfu/µL.
Each dilution has been tested in replicates (n = 4)
using Norgen’s Aspergillus niger PCR Detection Kit on
a 1X TAE 1.7% agarose gel.
The linear range of Norgen’s Aspergillus niger PCR
Detection Kit has been determined to cover
concentrations from 600 fg to 6 ng
Under the conditions of the Norgen’s Aspergillus
niger DNA Isolation procedure, Norgen’s Aspergillus
niger PCR Detection Kit covers a linear range from
100 copies to 1 x 105 copies/mg of plant tissue
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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Aspergillus niger PCR Detection Kit Cat. # 32900
Description Cat # Size
End-Point PCR 32900 24 rxns
Real-Time PCR TM32900 48 rxns
Real-Time PCR SG32900 48 rxns
Primer Sets and Controls 32910 100 rxns
Aspergillus niger Ordering information
Figure 1. Isolation and Detection of DNA from Fungi Samples with
No PCR Inhibition. Total genomic DNA was isolated from 40 mg
(wet weight) each of 8 economically important fungi species
affecting the grape industry using Norgen’s Fungi/Yeast
Genomic DNA Isolation Kit, which uses the same DNA isolation
technology as the Botytis cinerea PCR Detection Kit. The isolated
DNA was then subjected to quantitative PCR using universal
Intergenic Transcribed Space (ITS) gene primers to detect the
genomic DNA using the iQ SYBR Green Supermix (BioRad, #170-
8882). The black line below the PCR baseline is corresponding to
no template control while all other colored lines above the PCR
baseline are corresponding to the DNA isolated from 8 different
fungi species. The genomic DNA could be detected in all fungi
samples, indicating the high quality of the purified DNA.
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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A ready-to-use system for the isolation and
detection of B. cinerea using end-point PCR
Norgen’s Botrytis cinerea PCR Detection Kit is a ready-to-
use system for the isolation and detection of B.
cinerea from plant samples. First, the kit contains
components for the rapid isolation of total DNA, including
fungal DNA, from plant samples using spin-column
chromatography. Second, the kit contains B.
cinereaMaster Mix and controls to allow for PCR
amplification. The amplified PCR products are then
detected using agarose gel electrophoresis. Alternatively,
detection can be performed based on real-time PCR
using melt curves.
The B. cinerea Master Mix contains reagents and enzymes
for the specific amplification of a 381 bp region of the
fungal genome. In addition, Norgen’s B. cinerea PCR
Detection Kit contains a second Mastermix, the PCR
Control Master Mix, which can be used to identify possible
PCR inhibition and/or inadequate isolation via a separate
PCR reaction with the use of the PCR control (PCRC) or
Isolation Control (IsoC), respectively. This kit is designed to
allow for the testing of 24 samples. The Botrytis
cinerea PCR Primer Set and Controls are also available
separately for end-point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range of Norgen’s Botrytis cinerea PCR
Detection Kit was determined by analyzing a dilution
series of a B. cinerea quantification standards ranging
from 100 ag to 1 pg
Each dilution has been tested in replicates (n = 4)
using Norgen’s Botrytis cinerea PCR Detection Kit on a
1X TAE 1.7% agarose gel.
The linear range of Norgen’s Botrytis cinerea PCR
Detection Kit has been determined to cover
concentrations from 100 ag to 1 ng
Under the conditions of the Norgen’s Botrytis
cinerea DNA Isolation procedure, Norgen’s Botrytis
cinerea PCR Detection Kit covers a linear range from
100 copies to 1 x 106 copies.
Botrytis cinerea PCR Detection Kit Cat. # 29400
Description Cat # Size
End-Point PCR 29400 24 rxns
Real-Time PCR TM29400 48 rxns
Real-Time PCR SG29400 48 rxns
Primer Sets and Controls 29410 100 rxns
Botrytis cinerea Ordering information
Figure 1: Detection of C. cladosporioides using the C.
cladosporioides PCR Detection Kit. A representative 1X TAE 1.5%
agarose gel showing the amplification of serially diluted C.
cladosporioides positive (lane 1 to 4) negative (lane NTC)
controls. The size of the C. cladosporioides target amplicon
corresponds to 320 bp as represented by the provided DNA
Marker (M).
A ready-to-use system for the isolation and
detection of C.cladosporioides using end-point PCR
Norgen’s Cladosporium cladosporiodes PCR Detection Kit
is a ready-to-use system for the isolation and detection of
C. cladosporiodes from plant samples. First, the kit
contains components for the rapid isolation of total DNA,
including fungal DNA from plant samples using spin-
column chromatography based on Norgen’s proprietary
resin. Second, the kit contains C. cladosporiodes Master
Mix and controls to allow for PCR amplification, as well as
a Control Master Mix to allow for amplification of both an
Isolation Control and a PCR Control. The amplified PCR
products are then detected using agarose gel
electrophoresis. Alternatively, detection can be
performed based on real-time PCR using melt curves.
The C. cladosporioides Master Mix contains reagents and
enzymes for the specific amplification of a 320 bp region
o f t h e f u n g a l g e n o m e . I n a d d i t i o n ,
Norgen’s Cladosporium cladosporiodes PCR Detection Kit
contains a second Master Mix, the Control 2X PCR Master
Mix, which can be used to identify possible PCR inhibition
and/or inadequate isolation via a separate PCR with the
use of the provided PCR Control (PCRC) or Isolation
Control (IsoC). This kit is designed to allow for the testing of
24 samples. The Cladosporium cladosporiodes PCR Primer
Set and Controls are also available separately for end-
point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range was determined by analyzing a
dilution series of a C. cladosporioides quantification
standards ranging from 1 x 106 cfu/µL to 1 x 101 cfu/µL.
Each dilution has been tested in replicates (n = 4)
using on a 1X TAE 1.7% agarose gel.
The linear range of Norgen’s Cladosporium
cladosporioides PCR Detection Kit has been
determined to cover concentrations from 7 pg to 70
ng
Under the conditions of the
Norgen’s C.cladosporioides DNA Isolation procedure,
Norgen’s Cladosporium cladosporioides PCR
Detection Kit covers a linear range from 1500 copies
to 1.5 x 106 copies/50 mg of plant tissue
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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Cladosporium cladosporioides PCR Detection Kit Cat. # 33000
Description Cat # Size
End-Point PCR 33000 24 rxns
Real-Time PCR TM33000 48 rxns
Real-Time PCR SG33000 48 rxns
Primer Sets and Controls 33010 100 rxns
C. cladosporioides Ordering information
(320 bp)
Figure 1. Sensitivity of Detection using the Penicillium sp. PCR
Detection Kit. A representative 1X TAE, 1.7% agarose gel showing
the amplification of Penicillium sp. at different concentrations
(Target). The size of the Penicillium sp. target amplicon
corresponds to the 327 bp band represented by the provided
DNA Marker (M). The size of the Isolation Control corresponds to
the 550 bp band represented by the provided DNA Marker (M).
The Penicillium sp. 2X PCR Master Mix contains a PCR Control
which controls for PCR inhibition. The size of the PCR Control
corresponds to the 171 bp band represented by the provided
DNA Marker (M). Lanes A-E represents samples spiked with
different Penicillium sp. concentrations isolated from 50 mg plant
samples (interpreted as positive results). Lane F represents a no
fungus control.
Figure 2. Specificity of the Primers used to Detect Penicillium sp. A
representative 1X TAE, 1.7% agarose gel showing the specificity
of the primers used to detect the Penicillium sp. The size of
the Penicillium sp. target amplicon corresponds to the 327 bp
band represented by the provided DNA Marker (M). The
specificity of Norgen’s Penicillium sp. PCR Detection Kit is first and
foremost ensured by the selection of the Penicillium sp. specific
primers, as well as the selection of stringent reaction conditions.
The primers were checked for possible homologies in GenBank
published sequences by sequence comparison analyses.
Furthermore, the specificity of the Penicillium sp. specific primers
were tested against most of common fungi pathogens. Lane A
represents the gDNA from Aspergillus niger, Lane B represents the
gDNA from Cladosporium cladosporioides, Lane C represents the
gDNA from Botrytis cinerea, Lane D represents the gDNA
from Mucor racemosus, Lane E represents the gDNA
from Alternaria tenuissima, Lane F represents the gDNA
from Penicillin sp., Lane G represents the gDNA from Rhizopus
oryzae and Lane H represents the gDNA from Fusarium
oxysporum.
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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A ready-to-use system for the isolation and
detection of Penicillium sp. using end-point PCR
Norgen’s Penicillium sp. PCR Detection Kit is a ready-to-
use system for the isolation and detection of Penicillium
sp from plant samples. First, the kit contains components
for the rapid isolation of total DNA, including fungal DNA,
from plant samples using spin-column chromatography
based on Norgen’s proprietary resin. Second, the kit
contains Penicillium sp. Master Mix and controls to allow
for PCR amplification. The amplified PCR products are
then detected using agarose gel electrophoresis.
Alternatively, detection can be performed based on real-
time PCR using melt curves.
The Penicillium sp. Master Mix contains reagents and
enzymes for the specific amplification of a 327 bp region
of the fungal genome. In addition, Norgen’sPenicillium sp.
PCR Detection Kit contains a second heterologous
amplification system to identify possible PCR inhibition
and/or inadequate isolation. This kit is designed to allow
for the testing of 24 samples. The Penicillium sp PCR Primer
Set and Controls are also available separately for end-
point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range of Norgen’s Penicillium sp.PCR
Detection Kit was determined by analyzing a dilution
series of a Penicillium sp.quantification standards
ranging from 1 x 106 cfu/µL to 1 x 101 cfu/µL.
Each dilution has been tested in replicates (n = 4)
using Norgen’s Penicillium sp. PCR Detection Kit on a
1X TAE 1.7% agarose gel.
The linear range of Norgen’s Penicillium sp.PCR
Detection Kit has been determined to cover
concentrations from 18 pg to 18 ng
Under the conditions of the Norgen’s Penicillium
sp.DNA Isolation procedure, Norgen’s Penicillium
sp.PCR Detection Kit covers a linear range from 640
copies to 6.4 x 105 copies.
Penicillium sp. PCR Detection Kit Cat. # 33200
Description Cat # Size
End-Point PCR 33200 24 rxns
Real-Time PCR TM33200 48 rxns
Real-Time PCR SG33200 48 rxns
Primer Sets and Controls 33210 100 rxns
Penicillium sp. Ordering information
(327 bp)
Figure 1. Isolation and Detection of DNA from Yeast Samples with
No PCR Inhibition. Total genomic DNA was isolated from 500 µL
cultures of 4 economically important yeast species affecting the
grape industry using Norgen's Fungi/Yeast Genomic DNA
Isolation Kit, which uses the same DNA isolation technology as
the S. cerevisiae PCR Detection Kit. The isolated DNA was then
subjected to quantitative PCR using universal Intergenic
Transcribed Space (ITS) gene primers to detect the genomic DNA
using the iQ SYBR Green Supermix (BioRad, #170-8882). The black
line below the PCR baseline is corresponding to no template
control while all blue lines above the PCR baseline are
corresponding to the DNA isolated from 4 different yeast species.
The genomic DNA could be detected in all yeast samples,
indicating the high quality of the purified DNA.
Figure 2. Detection of S. cerevisiae using the S. cerevisiae PCR
Detection Kit. A representative 1X TAE 1.5% agarose gel showing
the amplification of S. cerevisiae positive (lane 1 and 2) negative
(lane 3 and 4) controls. The size of the S. cerevisiae target
amplicon corresponds to 301 bp as represented by the provided
DNA Marker (M).
A ready-to-use system for the isolation and
detection of S. cerevisiae using end-point PCR
Norgen’s Saccharomyces cerevisiae PCR Detection Kit is
a ready-to-use system for the isolation and detection of S.
cerevisiae from cultures, plants, and beverage samples.
First, the kit contains components for the rapid isolation of
total DNA, including fungal DNA, from thesamples using
spin-column chromatography based on Norgen’s
proprietary resin. Second, the kit contains S.
cerevisiae Master Mix and controls to allow for PCR
amplification. The amplified PCR products are then
detected using agarose gel electrophoresis. Alternatively,
detection can be performed based on real-time PCR
using melt curves.
The S. cerevisiae Master Mix contains reagents and
enzymes for the specific amplification of a 301 bp region
o f t h e f u n g a l g e n o m e . I n a d d i t i o n ,
Norgen’sSaccharomyces cerevisiae PCR Detection Kit
contains a second Mastermix, the PCR Control Master
Mix, which can be used to identify possible PCR inhibition
and/or inadequate isolation via the internal PCR control
(PCRC) or the provided Isolation Control (IsoC),
respectively. This kit is designed to allow for the testing of
24 samples. The Saccharomyces cerevisiae PCR Primer
Set and Controls are also available separately for end-
point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range of Norgen’s S. cerevisiae PCR
Detection Kit was determined by analyzing a dilution
series of a S. cerevisiae quantification standards
ranging from 1 x 106 cfu/µL to 1 x 101 cfu/µL.
Each dilution has been tested in replicates (n = 4)
using Norgen’s S. cerevisiaePCR Detection Kit on a 1X
TAE 1.7% agarose gel.
The linear range of Norgen’s S. cerevisiae PCR
Detection Kit has been determined to cover
concentrations from 110 pg to 11 ng
Under the conditions of the Norgen’s S.
cerevisiae DNA Isolation procedure, Norgen’s S.
cerevisiae PCR Detection Kit covers a linear range
from 800 copies to 8.5 x 105 copies.
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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Saccharomyces cerevisiae PCR Detection Kit Cat. # 33300
Description Cat # Size
End-Point PCR 33300 24 rxns
Real-Time PCR TM33300 48 rxns
Real-Time PCR SG33300 48 rxns
Primer Sets and Controls 33310 100 rxns
Saccharomyces cerevisiae Ordering information
Figure 1. Detection using the Plum Pox Virus RT-PCR Detection
Kit. A representative 1X TAE 1.5% agarose gel showing the
amplification of PPV negative (lanes 6 and 7) and PPV positive
(lanes 1 to 5) controls. The size of the PPV target amplicon
corresponds to 295 bp as represented by the provided DNA
Marker (M).
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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A ready-to-use system for the isolation and
detection of Plum Pox Virus using end-point PCR
Norgen’s Plum Pox Virus RT-PCR Detection Kit is a ready-to
-use system for the isolation and detection of Plum Pox
Virus from plant samples. First, the kit contains
components for the rapid isolation of total RNA, including
viral RNA, from plant samples using spin-column
chromatography based on Norgen’s proprietary resin.
Second, the kit contains Plum Pox Virus Master Mix and
controls to allow for PCR amplification, as well as a
Control Master Mix to allow for amplification of both an
isolation control and a PCR control. The amplified PCR
products are then detected using agarose gel
electrophoresis. Alternatively, detection can be
performed based on real-time PCR using melt curves.
The Plum Pox Virus Master Mix contains reagents and
enzymes for the specific amplification of a 295 bp region
of the viral genome. In addition, Norgen’s Plum Pox Virus
RT-PCR Detection Kit contains a second Mastermix, the
Control 2X RT-PCR Master Mix, which can be used to
identify possible PCR inhibition and/or inadequate
isolation via a separate RT-PCR reaction with the use of
the provided PCR control (PCRC) or Isolation Control
(IsoC), respectively. This kit is designed to allow for the
testing of 24 samples. The Plum Pox Virus RT-PCR Detection
Kit is also available in a 96-well format for high throughput
analysis. The Plum Pox Virus RT-PCR Primer Set and
Controls Controls are also available separately for end-
point PCR detection,
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range of Norgen’s PPV RT-PCR Detection Kit
was determined by analyzing a dilution series of an
PPV quantification standards ranging from 100 ag to 1
pg.
Each dilution has been tested in replicates (n = 4)
using Norgen’s PPV RT-PCR Detection Kit on a 1X TAE
1.5% agarose gel.
The linear range of Norgen’s PPV RT-PCR Detection Kit
has been determined to cover concentrations from
100 ag to 1 ng
Under the conditions of the Norgen’s PPV RNA
Isolation procedure, Norgen’s PPV RT-PCR Detection
Kit covers a linear range from 100 copies to 1 x 106
copies.
Plum Pox Virus PCR Detection Kit Cat. # 33400
Description Cat # Size
End-Point PCR 33400 24 rxns
Real-Time PCR TM33400 48 rxns
Real-Time PCR SG33400 48 rxns
Primer Sets and Controls 33410 100 rxns
Plum Pox Virus Ordering information
(295 bp)
Figure 1. Sensitivity of Detection using the Phomopsis viticola PCR
Detection Kit. A representative 1X TAE, 1.7% agarose gel showing
the amplification of P. viticola at different concentrations
(Target). The size of the P. viticola target amplicon corresponds to
the 300 bp band represented by the provided DNA Marker (M).
The size of the Isolation Control corresponds to the 529 bp band
represented by the provided DNA Marker (M). The P. viticola 2X
PCR Master Mix contains a PCR Control which controls for PCR
inhibition. The size of the PCR Control corresponds to the 171 bp
band represented by the provided DNA Marker (M). Lanes A-D
represents samples spiked with different P. viticola concentrations
isolated from 50 mg plant samples (interpreted as positive
results). Lane E represents a no fungus and Isolation control.
Figure 2. Specificity of the Primers used to Detect P. viticola. A
representative 1X TAE, 1.7% agarose gel showing the specificity
of the primers used to detect the P. viticola. The size of the P.
viticola target amplicon corresponds to the 300 bp band
represented by the provided DNA Marker (M). The specificity of
Norgen's P. viticola PCR Detection Kit is first and foremost ensured
by the selection of the P. viticola specific primers, as well as the
selection of stringent reaction conditions. The primers were
checked for possible homologies in GenBank published
sequences by sequence comparison analyses. Furthermore, the
specificity of the P. viticolaspecific primers were tested against
most of common fungi pathogens. Lane A represents the gDNA
from Aspergillus niger, Lane B represents the gDNA from
Cladosporium cladosporioides, Lane C represents the gDNA
from Botrytis cinerea, Lane D represents the gDNA from Mucor
racemosus, Lane E represents the gDNA from Alternaria
tenuissima, Lane F represents the gDNA from Penicillium sp., Lane
G represents the gDNA from Rhizopus oryzae, Lane H represents
the gDNA from Fusarium oxysporum and Lane I represents the
gDNA from P. viticola.
A ready-to-use system for the isolation and
detection of P. viticola using end-point PCR
Norgen’s Phomopsis viticola PCR Detection Kit is a ready-
to-use system for the isolation and detection of Phomopsis
viticola from plant samples. First, the kit contains
components for the rapid isolation of total DNA, including
fungal DNA, from plant samples using spin-column
chromatography based on Norgen’s proprietary resin.
Second, the kit contains Phomopsis viticola Master Mix
and controls to allow for PCR amplification. The amplified
PCR products are then detected using agarose gel
electrophoresis. Alternatively, detection can be
performed based on real-time PCR using melt curves.
The Phomopsis viticola. Master Mix contains reagents and
enzymes for the specific amplification of a 300 bp region
of the fungal genome. In addition, Norgen’s Phomopsis
viticola PCR Detection Kit contains a second
heterologous amplification system to identify possible PCR
inhibition and/or inadequate isolation. This kit is designed
to allow for the testing of 24 samples. ThePhomopsis
viticola PCR Primer Set and Controls are also available
separately for end-point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range (analytical measurement) of
Norgen’s Phomopsis viticola PCR Detection Kit was
determined by analyzing a dilution series of a P.
viticolaquantification standards ranging from 1 x
107 cfu/µL to 1 x 103 cfu/µL.
Each dilution has been tested in replicates (n = 4)
using Norgen’s Phomopsis viticola PCR Detection Kit
on a 1X TAE 1.7% agarose gel.
The linear range of Norgen’s Phomopsis viticola PCR
Detection Kit has been determined to cover
concentrations of genomic DNA from 10 pg to 100 ng
Under the conditions of the Norgen’s Phomopsis
viticola DNA Isolation procedure, Norgen’s Phomopsis
viticola PCR Detection Kit covers a linear range from
500 copies to 5 x 106 copies.
For research use only and NOT intended for in vitro diagnostics
Toll Free in North America: 1-866-667-4362 www.norgenbiotek.com
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Phomopsis viticola PCR Detection Kit Cat. # 34900
Description Cat # Size
End-Point PCR 34900 24 rxns
Real-Time PCR TM34900 48 rxns
Real-Time PCR SG34900 48 rxns
Primer Sets and Controls 34910 100 rxns
Phomopsis viticola Ordering information
(300 bp)
Figure 1. Sensitivity of Detection using the Erwinia amylovora PCR
Detection Kit. A representative 1X TAE 1.5% agarose gel showing
the amplification of Erwinia amylovora at different
concentrations. The size of theE. amylovora target amplicon
corresponds to 355 bp as represented by the provided DNA
Marker (M). NTC = Negative Control.
For research use only and NOT intended for in vitro diagnostics
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A ready-to-use system for the isolation and
detection of E. amylovora using end-point PCR
Norgen’s Erwinia amylovora PCR Detection Kit is a ready-
to-use system for the isolation and detection of Erwinia
amylovora from plant samples. First, the kit contains
components for the rapid isolation of total DNA, including
bacterial DNA, from bacterial cultures and plant samples
using spin-column chromatography based on Norgen’s
proprietary resin. Second, the kit contains Erwinia
amylovora Master Mix and controls to allow for PCR
amplification. The amplified PCR products are then
detected using agarose gel electrophoresis. Alternatively,
detection can be performed based on real-time PCR
using melt curves.
The Erwinia amylovora Master Mix contains reagents and
enzymes for the specific amplification of a 355 bp region
of the bacterial genome. In addition, Norgen’s Erwinia
amylovora PCR Detection Kit contains a second
heterologous amplification system to identify possible PCR
inhibition and/or inadequate isolation. This kit is designed
to allow for the testing of 24 samples. The Erwinia
amylovora PCR Primer Set and Controls are also available
separately for end-point PCR detection.
Features and Benefits
Rapid isolation of high quality DNA from plants
Contains two ready-to-use 2X PCR Master Mixes
High sensitivity and specificity
Includes an isolation control and a PCR control
Primer set and controls also available separately
Ideal for use in:
1. Field Surveillance of Pathogens
2. Surveys
Linear Range
The linear range (analytical measurement) of
Norgen’s Erwinia amylovora PCR Detection Kit was
determined by analyzing a dilution series of E.
amylovora quantification standards ranging from 1 x
106 cfu/µL to 1 x 102 cfu/µL.
Each dilution has been tested in replicates (n = 4)
using Norgen’s Erwinia amylovora PCR Detection Kit
on a 1X TAE 1.7% agarose gel.
The linear range of Norgen’s Erwinia amylovora PCR
Detection Kit has been determined to cover
concentrations of genomic DNA from 1 fg to 1 ng
Under the conditions of the Norgen’s Erwinia
amylovora DNA Isolation procedure, Norgen’s Erwinia
amylovora PCR Detection Kit covers a linear range
from 30 copies to 3 x 106 copies.
Erwinia amylovora PCR Detection Kit Cat. # 35100
Description Cat # Size
End-Point PCR 35100 24 rxns
Real-Time PCR TM35100 48 rxns
Real-Time PCR SG35100 48 rxns
Primer Sets and Controls 35110 100 rxns
Erwinia amylovora Ordering information
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Phone: (905) 227-8848
Toll Free: 1-866-NORGENB (667-4362)
Fax: (905) 227-1061
Email: [email protected]
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