GBI – Genera Biosystems Ltd.

Advanced Molecular Diagnostic Systems – Multiplex qPCR

Simultaneous quantification of the most common and proteolytic Pseudomonas species in raw milk by multiplex qPCR

Posted on by Bianca
Simultaneous quantification of the most common and proteolytic Pseudomonas species in raw milk by multiplex qPCR

The warmth-stable peptidase AprX, secreted by psychrotolerant Pseudomonas species in uncooked milk, is a serious explanation for destabilization and untimely spoilage of ultra-high temperature (UHT) milk and milk merchandise. To allow fast detection and quantification of seven frequent and proteolytic Pseudomonas species (P. proteolytica, P. gessardii, P. lactis, P. fluorescens, P. protegens, P. lundensis, and P. fragi) in uncooked milk, we developed two triplex qPCR assays bearing in mind species-dependent variations in AprX exercise.

Moreover 5 species-specific hydrolysis probes, focusing on the aprX gene, a common rpoB probe was included within the assay to find out the whole Pseudomonas counts. For all six probes, linear regression strains between Cq worth and goal DNA focus have been obtained in singleplex in addition to in multiplex approaches, yielding R2 values of > 0.975 and amplification efficiencies of 85-97%. Furthermore, excessive specificity was decided utilizing genomic DNA of 75 Pseudomonas strains, assigned to 57 species, and 40 different bacterial species as templates within the qPCR. Quantification of the goal species and whole Pseudomonas counts resulted in linear detection ranges of approx. 103-107 cfu/ml, which correspond properly to widespread Pseudomonas counts in uncooked milk.

Software of the assay utilizing 60 uncooked milk samples from completely different dairies confirmed good settlement of whole Pseudomonas counts calculated by qPCR with cell counts derived from cultivation. Moreover, a remarkably excessive variability relating to the species composition was noticed for every milk pattern, whereby P. lundensis and P. proteolytica/P. gessardii have been the predominant species detected. KEY POINTS: • Multiplex qPCR for quantification of seven proteolytic Pseudomonas species and whole Pseudomonas counts in uncooked milk • Excessive specificity and sensitivity through hydrolysis probes in opposition to aprX and rpoB • Fast methodology to find out Pseudomonas contamination in uncooked milk and predict spoilage potential.

Detection and Quantification of Rhizoctonia solani and Rhizoctonia solani AG1-IB Inflicting the Backside Rot of Lettuce in Tissues and Soils by Multiplex qPCR

Within the muck soil area of southwestern Quebec, vegetable growers are threatened by a number of soilborne illnesses, significantly the underside rot of lettuce brought on by the fungus Rhizoctonia solani. The very hot temperature of the few final seasons was marked by a rise in illness severity, and the related yield losses have been vital for Quebec lettuce growers. Within the absence of registered fungicides and resistant cultivars, the administration of Rhizoctonia solani-induced illnesses in lettuce is predicated on good agricultural practices, which require detailed data of the pathogen.

On this research, Rhizoctonia solani fungal strains have been remoted from contaminated field-grown lettuce vegetation presenting backside rot signs to find out the anastomotic teams (AGs) of those isolates by inside transcribed spacer area (ITS) sequencing. Rhizoctonia solani AG 1-IB was recognized as the primary anastomotic group inflicting backside rot lettuce in field-grown lettuce in natural soils within the Montérégie area. Two particular and delicate quantitative PCR assays have been then developed for R. solani AG1-IB and R. solani. The AG 1-IB qPCR assay amplified all strains of R. solani AG 1-IB examined, and no PCR product was obtained for any non-target strains.

Simultaneous quantification of the most common and proteolytic Pseudomonas species in raw milk by multiplex qPCR

The R. solani qPCR assay amplified all strains of R. solani and didn’t amplify non-target strains, besides for 2 strains of binucleate Rhizoctonia AG-E. In artificially inoculated soils, the sensitivity of each qPCR assays was set to 1 μg of sclerotia g-1 of dry soil. Within the development chamber experiment, a minimal focus between 14 and 42 μg sclerotia g-1 of dry soil was required to induce the event of signs on the lettuce. Certainly, the AG 1-IB qPCR assay was delicate sufficient to detect the bottom soil focus of AG1-IB able to inducing signs in head lettuce. As well as, the qPCR assays efficiently detected R. solani and R. solani AG 1-IB from contaminated plant tissue samples and soil samples from lettuce fields. The qPCR assays developed on this research might be helpful instruments in lettuce backside rot administration.

A delicate and reasonably priced multiplex RT-qPCR assay for SARS-CoV-2 detection

With the continuing COVID-19 (Coronavirus Illness 2019) pandemic, brought on by the novel coronavirus SARS-CoV-2 (Extreme Acute Respiratory Syndrome Coronavirus 2), there’s a want for delicate, particular, and reasonably priced diagnostic checks to determine contaminated people, not all of whom are symptomatic. Probably the most delicate take a look at includes the detection of viral RNA utilizing RT-qPCR (quantitative reverse transcription PCR), with many industrial kits now obtainable for this goal. Nonetheless, these are costly, and provide of such kits in enough numbers can’t at all times be assured.

We subsequently developed a multiplex assay utilizing well-established SARS-CoV-2 targets alongside a human mobile management (RPP30) and a viral spike-in management (Phocine Herpes Virus 1 [PhHV-1]), which monitor pattern high quality and nucleic acid extraction effectivity, respectively. Right here, we set up that this take a look at performs in addition to extensively used industrial assays, however at considerably diminished price. Moreover, we exhibit >1,000-fold variability in materials routinely collected by mixed nostril and throat swabbing and set up a statistically vital correlation between the detected stage of human and SARS-CoV-2 nucleic acids.

Pro QPCR SuperMix Kit
K5053200 200 reactions
EUR 188
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Pro QPCR SuperMix Kit
K5053400 400 reactions
EUR 299
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Green qPCR SuperMix
20-abx098031
  • EUR 1038.00
  • EUR 439.00
  • EUR 634.00
  • 15 ml
  • 1 ml
  • 5 ml
  • Shipped within 5-10 working days.

Probe qPCR SuperMix
20-abx098040
  • EUR 1038.00
  • EUR 439.00
  • EUR 634.00
  • 15 ml
  • 1 ml
  • 5 ml
  • Shipped within 5-10 working days.

Eva QPCR SuperMix Kit
K5052200 200 reactions
EUR 211
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Eva QPCR SuperMix Kit
K5052400 400 reactions
EUR 320
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Green qPCR SuperMix UDG
20-abx098032
  • EUR 1135.00
  • EUR 453.00
  • EUR 662.00
  • 15 ml
  • 1 ml
  • 5 ml
  • Shipped within 5-10 working days.

Top Green qPCR SuperMix
20-abx098033
  • EUR 829.00
  • EUR 217.00
  • EUR 1135.00
  • EUR 411.00
  • 15 ml
  • 1 ml
  • 25 ml
  • 5 ml
  • Shipped within 5-10 working days.

Tip Green qPCR SuperMix
20-abx098034
  • EUR 968.00
  • EUR 230.00
  • EUR 1330.00
  • EUR 467.00
  • 15 ml
  • 1 ml
  • 25 ml
  • 5 ml
  • Shipped within 5-10 working days.

Library Quantification qPCR SuperMix
20-abx098896
  • EUR 230.00
  • EUR 411.00
  • 1 ml
  • 5 ml
  • Shipped within 5-10 working days.

Power Eva QPCR SuperMix Kit
K5057200 200 reactions
EUR 188
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Power Eva QPCR SuperMix Kit
K5057400 400 reactions
EUR 320
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

cDNA Synthesis SuperMix for qPCR
20-abx09801920ulSystems
  • EUR 690.00
  • EUR 565.00
  • 100 rxns ×20 ul Systems
  • 50 rxns × 20 ul Systems
  • Shipped within 5-10 working days.

Top Green qPCR SuperMix (+Dye II)
20-abx098890
  • EUR 829.00
  • EUR 217.00
  • EUR 1135.00
  • EUR 411.00
  • 15 ml
  • 1 ml
  • 25 ml
  • 5 ml
  • Shipped within 5-10 working days.

Tip Green qPCR SuperMix (+Dye II)
20-abx098891
  • EUR 968.00
  • EUR 230.00
  • EUR 1330.00
  • EUR 467.00
  • 15 ml
  • 1 ml
  • 25 ml
  • 5 ml
  • Shipped within 5-10 working days.

HotTaq Probe qPCR Mix (ROX)
BT11001 250rxn
EUR 109
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq EvaGreen qPCR Mix (ROX)
BT11101 250rxn
EUR 123
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

TaqProbe 5X qPCR MasterMix-ROX
MasterMix-5P 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

BrightGreen 5X qPCR MasterMix-ROX
MasterMix-5R 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

KiloGreen 2X qPCR MasterMix-ROX
MasterMix-KR 4 x 1.25 ml - 500 reactions (20 ul)
EUR 140

BrightGreen miRNA qPCR MasterMix-ROX
MasterMix-mR 4 x 1.25 ml for 500 reactions
EUR 151

TaqProbe 2X qPCR MasterMix-ROX
MasterMix-P 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-ROX
MasterMix-R 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-ROX
MasterMix-R-XL 16 x 1.25 ml for 2000 reactions (20 ul)
EUR 376

VAHTS Library Quantification Kit for Illumina (Low ROX Premixed)
NQ103 500 rxn
EUR 291

VAHTS Library Quantification Kit for Illumina (High ROX Premixed)
NQ104 500 rxn
EUR 291

cDNA Synthesis SuperMix for qPCR (GC-rich)
abx098024-50rxns20ulSystems 50 rxns × 20 ul Systems
EUR 718
  • Shipped within 5-10 working days.

HiScript II Q RT SuperMix for qPCR
R222-01 100 rxn (20 μl/rxn)
EUR 235

HotTaq Probe qPCR Mix (no ROX)
BT11002 250rxn
EUR 109
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq EvaGreen qPCR Mix (no ROX)
BT11102 250rxn
EUR 123
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

BrightGreen 5X qPCR MasterMix-Low ROX
MasterMix-5L 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

TaqProbe 5X qPCR MasterMix-Low ROX
MasterMix-5PL 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

BrightGreen Express 2X qPCR MasterMix-ROX
MasterMix-ER 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

KiloGreen 2X qPCR MasterMix-Low ROX
MasterMix-KL 4 x 1.25 ml - 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-Low ROX
MasterMix-LR 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-Low ROX
MasterMix-LR-XL 16 x 1.25 ml for 2000 reactions (20 ul)
EUR 376

BrightGreen miRNA qPCR MasterMix-Low ROX
MasterMix-mL 4 x 1.25 ml for 500 reactions
EUR 151

TaqProbe 2X qPCR MasterMix-Low ROX
MasterMix-PL 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

HiScript II Q Select RT SuperMix for qPCR
R232-01 100 rxn (20 μl/rxn)
EUR 235

ROX reference dye for qPCR, 500 uL
31110 500 uL
EUR 46

ROX reference dye for qPCR, 1 mL
41110 1 mL
EUR 52

SYBR Green qPCR Master Mix (High ROX)
HY-K0521 1 mL (100 rxns)
EUR 113

SYBR Green qPCR Master Mix (Low ROX)
HY-K0522 5 mL (500 rxns )
EUR 257

SYBR Green qPCR Master Mix (No ROX)
HY-K0523 5 mL (500 rxns )
EUR 257

BrightGreen Express 2X qPCR MasterMix-Low ROX
MasterMix-EL 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

Accuris qMax Probe No Rox qPCR Mix
PR2001-N-1000 1 PC
EUR 467.74
  • To order instruments in 115V / US plug please delete the 'E' off the order code.European 2 pin plugs will be supplied as standard, please request UK if required.

HiScript III RT SuperMix for qPCR (+g DNA wiper)
R323-01 100 rxn (20 μl/rxn)
EUR 293

PLATEMAX ULTRA CLEAR PERMANENT HEAT SEALING FILM FOR QPCR, 100/500
HS-100-QPCR 100/pk
EUR 318
Description: Sealing Products; Sealing films - Axygen

PLATEMAX ULTRA CLEAR PEELABLE HEAT SEALING FILM FOR QPCR. 100/500
HS-150-QPCR 100/pk
EUR 318
Description: Sealing Products; Sealing films - Axygen

2x SYBR Green qPCR Master Mix (High ROX)
B21402 5 ml
EUR 224
Description: 2x SYBR Green qPCR master mix (High ROX) utilizes a special performance-enhanced Taq DNA polymerase protected via a hot-start activation technique, and optimized qPCR buffer system to perform SYBR

2x SYBR Green qPCR Master Mix (High ROX)
B21403 25 ml
EUR 856
Description: 2x SYBR Green qPCR master mix (High ROX) utilizes a special performance-enhanced Taq DNA polymerase protected via a hot-start activation technique, and optimized qPCR buffer system to perform SYBR

2x SYBR Green qPCR Master Mix (Low ROX)
B21702 5 ml
EUR 224
Description: 2x SYBR Green qPCR master mix (Low ROX) utilizes a special performance-enhanced Taq DNA polymerase protected via a hot-start activation technique, and optimized qPCR buffer system to perform SYBR

2x SYBR Green qPCR Master Mix (Low ROX)
B21703 25 ml
EUR 856
Description: 2x SYBR Green qPCR master mix (Low ROX) utilizes a special performance-enhanced Taq DNA polymerase protected via a hot-start activation technique, and optimized qPCR buffer system to perform SYBR

Green-2-Go qPCR Mastermix-ROX (500X20ul Rxn)
QPCR001-R 4X1.25ml
EUR 252.71
  • Product category: PCR Related/PCR Premix (qPCR)/qPCR Premix + Taq

QCell-Pro One-Step qRT-PCR SuperMix Kit
K5055200 200 reactions
EUR 376
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

QCell-Pro One-Step qRT-PCR SuperMix Kit
K5055400 400 reactions
EUR 572
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

AzuraView GreenFast qPCR Blue Mix Lo Rox - 200 Rxn
AZ-2301 200 Rxn
EUR 179

AzuraView GreenFast qPCR Blue Mix Lo Rox - 500 Rxn
AZ-2305 500 Rxn
EUR 320

AzuraView GreenFast qPCR Blue Mix Lo Rox - 2000 Rxn
AZ-2320 2000 Rxn
EUR 996

AzuraView GreenFast qPCR Blue Mix Lo Rox - 5000 Rxn
AZ-2325 5000 Rxn
EUR 2348

AzuraView GreenFast qPCR Blue Mix Hi Rox - 200 Rxn
AZ-2401 200 Rxn
EUR 179

AzuraView GreenFast qPCR Blue Mix Hi Rox - 500 Rxn
AZ-2405 500 Rxn
EUR 320

AzuraView GreenFast qPCR Blue Mix Hi Rox - 2000 Rxn
AZ-2420 2000 Rxn
EUR 996

AzuraView GreenFast qPCR Blue Mix Hi Rox - 5000 Rxn
AZ-2421 5000 Rxn
EUR 2348

Green-2-Go qPCR Mastermix-Low ROX (500X20ul Rxn)
QPCR002-L 4X1.25ml
EUR 252.71
  • Product category: PCR Related/PCR Premix (qPCR)/qPCR Premix + Taq

HiScript II Q Select RT SuperMix for qPCR (+g DNA wiper)
R233-01 100 rxn (20 μl/rxn)
EUR 244

Ultra SYBR Green qPCR Master Mix (2X, with ROX I)
W2601-1 NULL
EUR 0

Ultra SYBR Green qPCR Master Mix (2X, with ROX I)
W2601-5 NULL
EUR 0

T-Pro BCA Protein Assay kit
JB04-D001 500assay/KIT
EUR 204

T-Pro Plasmid Mini Kit (100)
RB94-YPD100 100preps/Kit
EUR 161

T-Pro Plasmid Mini Kit (250)
RB94-YPD250 250preps/Kit
EUR 222

T-Pro Plasmid Midi Kit (20)
RB94-YPI020 20preps/Kit
EUR 213

T-Pro Plasmid Maxi Kit (10)
RB94-YPM010 10preps/Kit
EUR 222

Fast Plus EvaGreen Master Mix for qPCR with Rox (200 rxn)
31014 2x1mL
EUR 228
Description: Minimum order quantity: 1 unit of 2x1mL

Fast Plus EvaGreen qPCR Master Mix with low Rox (500 rxn)
31014-1 5x1mL
EUR 437
Description: Minimum order quantity: 1 unit of 5x1mL

Fast Plus EvaGreen qPCR Master Mix with low Rox (5000 rxn)
31014-2 50x1mL
EUR 3576
Description: Minimum order quantity: 1 unit of 50x1mL

Fast Plus EvaGreen qPCR Master Mix with high Rox (500 rxn)
31015-1 5x1mL
EUR 437
Description: Minimum order quantity: 1 unit of 5x1mL

Fast Plus EvaGreen qPCR Master Mix with high Rox (5000 rxn)
31015-2 50x1mL
EUR 3576
Description: Minimum order quantity: 1 unit of 50x1mL

Forget-Me-Notâ„¢ EvaGreen qPCR Master Mix with ROX (500 reactions)
31042-1 5x1mL
EUR 272
Description: Minimum order quantity: 1 unit of 5x1mL

Forget-Me-Notâ„¢ EvaGreen qPCR Master Mix with ROX (200 reactions)
31042-20mL 2x10mL
EUR 494
Description: Minimum order quantity: 1 unit of 2x10mL

Forget-Me-Notâ„¢ EvaGreen qPCR Master Mix with ROX (100 reactions)
31042-T 1mL
EUR 114
Description: Minimum order quantity: 1 unit of 1mL

Forget-Me-Notâ„¢ EvaGreen qPCR Master Mix (Low ROX) (500 reactions)
9-31045
  • EUR 114.00
  • EUR 482.00
  • EUR 266.00
  • 1mL
  • 1X10mL
  • 5x1mL
Description: Minimum order quantity: 1 unit of 5x1mL

Forget-Me-Notâ„¢ EvaGreen qPCR Master Mix (High ROX) (500 reactions)
9-31046
  • EUR 114.00
  • EUR 482.00
  • EUR 266.00
  • 1mL
  • 1X10mL
  • 5x1mL
Description: Minimum order quantity: 1 unit of 5x1mL

PCR SuperMix
20-abx098887
  • EUR 272.00
  • EUR 578.00
  • 1 ml
  • 5 ml
  • Shipped within 5-10 working days.

T-Pro Stacking Buffer
JB03-C002 500ml/BT
EUR 152

T-Pro MycoClean spray
JT90-R002 500ml/BT
EUR 144

T-Pro Bradford Protein Assay kit(1X)
JB04-D002 500ml/BT
EUR 161

T-Pro Genomic DNA Midi Kit (20)
RB94-NGM020 20preps/kit
EUR 222

T-Pro Genomic DNA Mini Kit (100)
RB94-NGS100 100preps/kit
EUR 213

T-Pro LumiFast Plus Chemiluminescence Detection Kit (ECL Kit)
JT96-K002M 250ml*2/Kit
EUR 204

T-Pro LumiFast Plus Chemiluminescence Detection Kit (ECL Kit)
JT96-K002S 100ml*2/Kit
EUR 152

T-Pro LumiLong Plus Chemiluminescence Detection Kit (ECL Kit)
JT96-K004M 250ml*2/Kit
EUR 239

T-Pro LumiLong Plus Chemiluminescence Detection Kit (ECL Kit)
JT96-K004S 100ml*2/Kit
EUR 170

T-Pro Endotoxin Removal Plasmid Midi kit (25)
RB94-EPI020 20preps/Kit
EUR 248

T-Pro Endotoxin Removal Plasmid Maxi kit (10)
RB94-EPM010 10preps/Kit
EUR 265

T-Pro Plant Genomic DNA Midi Kit (20)
RB94-PGM020 20preps/kit
EUR 222

The inclusion of the human management probe in our assay subsequently supplies a quantitative measure of pattern high quality that would assist cut back false-negative charges. We exhibit the feasibility of building a sturdy RT-qPCR assay at roughly 10% of the price of equal industrial assays, which may gain advantage low-resource environments and make high-volume testing reasonably priced.

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