The power to detect and quantify aerially dispersed plant pathogens is crucial for growing efficient illness management measures and epidemiological fashions that optimize the timing for management . There’s an acute want for managing the downy mildew pathogens infecting cucurbits and hop incited by members of the genus Pseudoperonospora (P. cubensis clade 1 and a couple of isolates and P. humuli, respectively). A extremely particular multiplex TaqMan qPCR assay concentrating on distinctive sequences within the pathogens’ mitochondrial genomes was developed that permits detection of all three taxa in a single multiplexed amplification.
An inner management included within the response evaluated if outcomes have been influenced by PCR inhibitors that may make it by way of the DNA extraction course of. Dependable quantification of inoculum as little as three sporangia in a pattern was noticed. The multiplexed assay was examined with DNA extracted from purified sporangia, contaminated plant tissue and environmental samples collected on impaction spore traps samplers. The power to precisely detect and concurrently quantify all three pathogens in a single multiplexed amplification ought to enhance administration choices for controlling the ailments they trigger.
A selected area of Cm (clvG gene) was chosen for primer design utilizing comparative genomics strategy. A completely artificial common inner management (UIC) was additionally designed to detect PCR inhibitors and false detrimental leads to qPCR reactions. The Cm primers can be utilized alone or in a triplex TaqMan qPCR assay with UIC and beforehand described Clavibacter-specific primers. The assay was particular for Cm and detected upto 10 fg of Cm DNA in sensitivity and spiked assay. Addition of the UIC didn’t change the specificity or sensitivity of the multiplex TaqMan qPCR assay.
Particular and Delicate Detection of Neuroblastoma mRNA Markers by Multiplex RT-qPCR
mRNA RT-qPCR is proven to be a really delicate method to detect minimal residual illness (MRD) in sufferers with neuroblastoma. A number of mRNA markers are identified to detect heterogeneous neuroblastoma cells in bone marrow (BM) or blood from sufferers. Nevertheless, the restricted volumes of BM and blood obtainable can hamper the detection of a number of markers. To make optimum use of those samples, we developed a multiplex RT-qPCR for the detection of MRD in neuroblastoma. GUSB and PHOX2B have been examined as single markers. The adrenergic markers TH, GAP43, CHRNA3 and DBH and mesenchymal markers POSTN, PRRX1 and FMO3 have been examined in multiplex. Utilizing management blood and BM, we established new thresholds for positivity.
Comparability of multiplex and singleplex RT-qPCR outcomes from 21 blood and 24 BM samples from neuroblastoma sufferers demonstrated a comparable sensitivity. With this multiplex RT-qPCR, we’re capable of check seven totally different neuroblastoma mRNA markers, which overcomes tumor heterogeneity and improves sensitivity of MRD detection, even in these samples of low RNA amount. With sources and time being saved, discount in pattern quantity and consumables can help within the introduction of MRD by RT-qPCR into scientific follow. The outcomes demonstrated the potential of quantifying as few as 2.5 copies/response and evaluation of 244 sufferers with identified outcomes chosen by cross-section revealed 100% settlement with a qualitative RT-qPCR assay registered by Anvisa. On this inhabitants, it was doable to quantify sufferers with between 2.59 and three.5 × 107 copies per response and detrimental sufferers continued to point the identical end result.
The MRD disk: automated minimal residual illness monitoring by extremely delicate centrifugal microfluidic multiplex qPCR
We current a proof-of-principle research on automated, extremely delicate and multiplexed qPCR quantification by centrifugal microfluidics. The MRD disk can be utilized for standardisation of repetitive, longitudinal assays with excessive necessities on reproducibility and sensitivity, equivalent to most cancers monitoring. In distinction to high-throughput qPCR automation by cumbersome and costly robotic workstations we make use of a small centrifugal microfluidic instrument, addressing the necessity of low- to mid-throughput purposes. As a possible utility we display automated minimal residual illness (MRD) monitoring of prognostic markers in sufferers with acute lymphoblastic leukaemia (ALL).
The disk-workflow covers all features of scientific gold customary MRD quantification: era of normal curves, specificity controls, no template controls and quantification of the ALL affected person pattern. We built-in a extremely delicate, colorimetric 2-plex evaluation of MRD targets, in addition to a 2-plex evaluation of reference genes, each in parallel and in a single LabDisk cartridge. For this function, a scientific process for crosstalk- and signal-to-noise-optimisation is launched, offering a suggestion for environment friendly multiplex readout inside microfluidic platforms. The qPCR customary curves (n = 12/12) generated on-disk attain clinically required linearity (R2 = 98.1% to R2 = 99.8%).
In three consecutive MRD disk runs with an ALL affected person pattern containing the 2 consultant MRD targets VH3D3D5JH3 and VkIkde, we observe excessive accordance between the on-disk quantifications (48 ± 6 copies/response and 69 ± 6 copies/response) and the anticipated concentrations (57 copies/response for each targets). Compared to the scientific gold customary of manually pipetted, singleplex assays, the MRD disk yields comparable restrict of quantification (1 × 10-4) in n = 6/6 analyses (vs. n = 4/Four in gold customary) and a restrict of detection (1 × 10-5) in n = 6/6 evaluation (vs. n = 2/Four in gold customary).
 Green qPCR SuperMix |
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20-abx098031 |
Abbexa |
-
EUR 1038.00
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EUR 439.00
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EUR 634.00
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 Probe qPCR SuperMix |
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20-abx098040 |
Abbexa |
-
EUR 1038.00
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EUR 439.00
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EUR 634.00
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) Genious 2X SYBRGREEN FAST qPCR MIX (Low ROX) |
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RK21206 |
Abclonal |
1 ml |
Ask for price |
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 Power Eva QPCR SuperMix Kit |
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K5057200 |
Biochain |
200 reactions |
EUR 188 |
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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 |
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K5057400 |
Biochain |
400 reactions |
EUR 320 |
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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 |
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20-abx098032 |
Abbexa |
-
EUR 1135.00
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EUR 453.00
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EUR 662.00
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 Top Green qPCR SuperMix |
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20-abx098033 |
Abbexa |
-
EUR 829.00
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EUR 217.00
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EUR 1135.00
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EUR 411.00
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 Tip Green qPCR SuperMix |
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20-abx098034 |
Abbexa |
-
EUR 968.00
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EUR 230.00
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EUR 1330.00
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EUR 467.00
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 Library Quantification qPCR SuperMix |
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20-abx098896 |
Abbexa |
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) Genious 2X SYBR GREEN FAST qPCR MIX (No ROX) |
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RK21205 |
Abclonal |
1 ml |
Ask for price |
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 cDNA Synthesis SuperMix for qPCR |
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20-abx09801920ulSystems |
Abbexa |
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- 100 rxns ×20 ul Systems
- 50 rxns × 20 ul Systems
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 Methylamp MS-qPCR Fast Kit |
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P-1028 |
EpiGentek |
200 Reactions |
EUR 496.6 |
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Description: fast delivery possible |
) Fast Plus EvaGreen Master Mix for qPCR with Rox (200 rxn) |
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31014 |
Biotium |
2x1mL |
EUR 228 |
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Description: Minimum order quantity: 1 unit of 2x1mL |
) Fast Plus EvaGreen qPCR Master Mix with low Rox (500 rxn) |
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31014-1 |
Biotium |
5x1mL |
EUR 437 |
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Description: Minimum order quantity: 1 unit of 5x1mL |
) Fast Plus EvaGreen qPCR Master Mix with low Rox (5000 rxn) |
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31014-2 |
Biotium |
50x1mL |
EUR 3576 |
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Description: Minimum order quantity: 1 unit of 50x1mL |
) Fast Plus EvaGreen qPCR Master Mix with high Rox (500 rxn) |
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31015-1 |
Biotium |
5x1mL |
EUR 437 |
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Description: Minimum order quantity: 1 unit of 5x1mL |
) Fast Plus EvaGreen qPCR Master Mix with high Rox (5000 rxn) |
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31015-2 |
Biotium |
50x1mL |
EUR 3576 |
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Description: Minimum order quantity: 1 unit of 50x1mL |
) Top Green qPCR SuperMix (+Dye II) |
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20-abx098890 |
Abbexa |
-
EUR 829.00
-
EUR 217.00
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EUR 1135.00
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EUR 411.00
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) Tip Green qPCR SuperMix (+Dye II) |
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20-abx098891 |
Abbexa |
-
EUR 968.00
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EUR 230.00
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EUR 1330.00
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EUR 467.00
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) VAHTS Library Quantification Kit for Illumina (Low ROX Premixed) |
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NQ103 |
Vazyme |
500 rxn |
EUR 291 |
) VAHTS Library Quantification Kit for Illumina (High ROX Premixed) |
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NQ104 |
Vazyme |
500 rxn |
EUR 291 |
) HotTaq Probe qPCR Mix (ROX) |
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BT11001 |
Bioatlas |
250rxn |
EUR 109 |
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Description: High quality HotTaq polymerase for different PCR variations and downstream applications. |
) HotTaq EvaGreen qPCR Mix (ROX) |
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BT11101 |
Bioatlas |
250rxn |
EUR 123 |
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Description: High quality HotTaq polymerase for different PCR variations and downstream applications. |
 TaqProbe 5X qPCR MasterMix-ROX |
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MasterMix-5P |
ABM |
4 x 1.0 ml for 1000 reactions (20 ul) |
EUR 213 |
 BrightGreen 5X qPCR MasterMix-ROX |
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MasterMix-5R |
ABM |
4 x 1.0 ml for 1000 reactions (20 ul) |
EUR 213 |
 KiloGreen 2X qPCR MasterMix-ROX |
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MasterMix-KR |
ABM |
4 x 1.25 ml - 500 reactions (20 ul) |
EUR 140 |
 BrightGreen miRNA qPCR MasterMix-ROX |
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MasterMix-mR |
ABM |
4 x 1.25 ml for 500 reactions |
EUR 151 |
 TaqProbe 2X qPCR MasterMix-ROX |
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MasterMix-P |
ABM |
4 x 1.25 ml for 500 reactions (20 ul) |
EUR 140 |
 BrightGreen 2X qPCR MasterMix-ROX |
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MasterMix-R |
ABM |
4 x 1.25 ml for 500 reactions (20 ul) |
EUR 140 |
BrightGreen 2X qPCR MasterMix-ROX |
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MasterMix-R-XL |
ABM |
16 x 1.25 ml for 2000 reactions (20 ul) |
EUR 376 |
: (1mL)) Fast Plus EvaGreen qPCR Master Mix with low Rox (100 rxn): (1mL) |
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31014-T |
Biotium |
1mL |
EUR 134 |
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Description: Minimum order quantity: 1 unit of 1mL |
) Fast Plus EvaGreen Master Mix for qPCR with High Rox (200 rxn) |
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31015 |
Biotium |
2x1mL |
EUR 228 |
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Description: Minimum order quantity: 1 unit of 2x1mL |
: (1mL)) Fast Plus EvaGreen qPCR Master Mix with High Rox (100 rxn): (1mL) |
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31015-T |
Biotium |
1mL |
EUR 134 |
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Description: Minimum order quantity: 1 unit of 1mL |
) cDNA Synthesis SuperMix for qPCR (GC-rich) |
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abx098024-50rxns20ulSystems |
Abbexa |
50 rxns × 20 ul Systems |
EUR 718 |
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 HiScript II Q RT SuperMix for qPCR |
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R222-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 235 |
) HotTaq Probe qPCR Mix (no ROX) |
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BT11002 |
Bioatlas |
250rxn |
EUR 109 |
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Description: High quality HotTaq polymerase for different PCR variations and downstream applications. |
) HotTaq EvaGreen qPCR Mix (no ROX) |
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BT11102 |
Bioatlas |
250rxn |
EUR 123 |
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Description: High quality HotTaq polymerase for different PCR variations and downstream applications. |
 BrightGreen 5X qPCR MasterMix-Low ROX |
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MasterMix-5L |
ABM |
4 x 1.0 ml for 1000 reactions (20 ul) |
EUR 213 |
 TaqProbe 5X qPCR MasterMix-Low ROX |
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MasterMix-5PL |
ABM |
4 x 1.0 ml for 1000 reactions (20 ul) |
EUR 213 |
 BrightGreen Express 2X qPCR MasterMix-ROX |
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MasterMix-ER |
ABM |
4 x 1.25 ml for 500 reactions (20 ul) |
EUR 140 |
 KiloGreen 2X qPCR MasterMix-Low ROX |
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MasterMix-KL |
ABM |
4 x 1.25 ml - 500 reactions (20 ul) |
EUR 140 |
 BrightGreen 2X qPCR MasterMix-Low ROX |
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MasterMix-LR |
ABM |
4 x 1.25 ml for 500 reactions (20 ul) |
EUR 140 |
BrightGreen 2X qPCR MasterMix-Low ROX |
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MasterMix-LR-XL |
ABM |
16 x 1.25 ml for 2000 reactions (20 ul) |
EUR 376 |
 BrightGreen miRNA qPCR MasterMix-Low ROX |
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MasterMix-mL |
ABM |
4 x 1.25 ml for 500 reactions |
EUR 151 |
 TaqProbe 2X qPCR MasterMix-Low ROX |
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MasterMix-PL |
ABM |
4 x 1.25 ml for 500 reactions (20 ul) |
EUR 140 |
 HiScript II Q Select RT SuperMix for qPCR |
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R232-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 235 |
) SYBR Green qPCR Master Mix (High ROX) |
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HY-K0521 |
MedChemExpress |
1 mL (100 rxns) |
EUR 113 |
) SYBR Green qPCR Master Mix (Low ROX) |
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HY-K0522 |
MedChemExpress |
5 mL (500 rxns ) |
EUR 257 |
) SYBR Green qPCR Master Mix (No ROX) |
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HY-K0523 |
MedChemExpress |
5 mL (500 rxns ) |
EUR 257 |
 BrightGreen Express 2X qPCR MasterMix-Low ROX |
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MasterMix-EL |
ABM |
4 x 1.25 ml for 500 reactions (20 ul) |
EUR 140 |
 ROX reference dye for qPCR, 1 mL |
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41110 |
Lumiprobe |
1 mL |
EUR 52 |
 ROX reference dye for qPCR, 500 uL |
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31110 |
Lumiprobe |
500 uL |
EUR 46 |
) HiScript III RT SuperMix for qPCR (+g DNA wiper) |
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R323-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 293 |
 PLATEMAX ULTRA CLEAR PERMANENT HEAT SEALING FILM FOR QPCR, 100/500 |
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HS-100-QPCR |
CORNING |
100/pk |
EUR 318 |
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Description: Sealing Products; Sealing films - Axygen |
 PLATEMAX ULTRA CLEAR PEELABLE HEAT SEALING FILM FOR QPCR. 100/500 |
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HS-150-QPCR |
CORNING |
100/pk |
EUR 318 |
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Description: Sealing Products; Sealing films - Axygen |
 2X Universal SYBR Green Fast qPCR Mix |
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RK21203 |
Abclonal |
1 mL |
Ask for price |
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 Genious 2X SYBR Green Fast qPCR Mix |
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RK21204 |
Abclonal |
1 mL |
Ask for price |
|
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 QCell-Pro One-Step qRT-PCR SuperMix Kit |
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K5055200 |
Biochain |
200 reactions |
EUR 376 |
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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 |
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K5055400 |
Biochain |
400 reactions |
EUR 572 |
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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-2-Go qPCR Mastermix-ROX (500X20ul Rxn) |
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QPCR001-R |
Bio Basic |
4X1.25ml |
EUR 252.71 |
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) 2x SYBR Green qPCR Master Mix (High ROX) |
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B21402 |
Bimake |
5 ml |
EUR 224 |
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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) |
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B21403 |
Bimake |
25 ml |
EUR 856 |
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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) |
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B21702 |
Bimake |
5 ml |
EUR 224 |
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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) |
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B21703 |
Bimake |
25 ml |
EUR 856 |
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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 |
 and Fast ligase (2.5ul)]) Cas9 SmartNuclease Extra Ligation Kit [includes 5x ligation buffer (10 ul) and Fast ligase (2.5ul)] |
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CAS9LIG-KIT |
SBI |
1 Kit |
EUR 153 |
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) Fast EvaGreen Master Mix for qPCR(200 rxn) |
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31003 |
Biotium |
2x1ML |
EUR 204 |
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Description: Minimum order quantity: 1 unit of 2x1ML |
) Fast EvaGreen Master Mix for qPCR (500 rxn) |
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31003-1 |
Biotium |
5x1ML |
EUR 386 |
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Description: Minimum order quantity: 1 unit of 5x1ML |
) Fast EvaGreen Master Mix for qPCR (5000 rxn) |
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31003-2 |
Biotium |
50x1ML |
EUR 3116 |
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Description: Minimum order quantity: 1 unit of 50x1ML |
) Fast Plus EvaGreen qPCR Master Mix (200 rxn) |
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31020 |
Biotium |
2x1mL |
EUR 228 |
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Description: Minimum order quantity: 1 unit of 2x1mL |
) Fast Plus EvaGreen qPCR Master Mix (500 rxn) |
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31020-1 |
Biotium |
5x1mL |
EUR 437 |
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Description: Minimum order quantity: 1 unit of 5x1mL |
) Fast Plus EvaGreen qPCR Master Mix (5000 rxn) |
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31020-2 |
Biotium |
50x1mL |
EUR 3576 |
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Description: Minimum order quantity: 1 unit of 50x1mL |
) Green-2-Go qPCR Mastermix-Low ROX (500X20ul Rxn) |
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QPCR002-L |
Bio Basic |
4X1.25ml |
EUR 252.71 |
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 AzuraView GreenFast qPCR Blue Mix Lo Rox - 200 Rxn |
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AZ-2301 |
Real Time Primers |
200 Rxn |
EUR 179 |
 AzuraView GreenFast qPCR Blue Mix Lo Rox - 500 Rxn |
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AZ-2305 |
Real Time Primers |
500 Rxn |
EUR 320 |
 AzuraView GreenFast qPCR Blue Mix Lo Rox - 2000 Rxn |
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AZ-2320 |
Real Time Primers |
2000 Rxn |
EUR 996 |
 AzuraView GreenFast qPCR Blue Mix Lo Rox - 5000 Rxn |
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AZ-2325 |
Real Time Primers |
5000 Rxn |
EUR 2348 |
 AzuraView GreenFast qPCR Blue Mix Hi Rox - 200 Rxn |
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AZ-2401 |
Real Time Primers |
200 Rxn |
EUR 179 |
 AzuraView GreenFast qPCR Blue Mix Hi Rox - 500 Rxn |
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AZ-2405 |
Real Time Primers |
500 Rxn |
EUR 320 |
 AzuraView GreenFast qPCR Blue Mix Hi Rox - 2000 Rxn |
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AZ-2420 |
Real Time Primers |
2000 Rxn |
EUR 996 |
 AzuraView GreenFast qPCR Blue Mix Hi Rox - 5000 Rxn |
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AZ-2421 |
Real Time Primers |
5000 Rxn |
EUR 2348 |
) Fast Probe Master Mix with Rox (200 rxn) |
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31016 |
Biotium |
2x1mL |
EUR 234 |
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Description: Minimum order quantity: 1 unit of 2x1mL |
) Fast Probe Master Mix with Rox (500 rxn) |
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31016-1 |
Biotium |
5x1mL |
EUR 466 |
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Description: Minimum order quantity: 1 unit of 5x1mL |
) Fast Probe Master Mix with Rox (5000 rxn) |
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31016-2 |
Biotium |
50x1mL |
EUR 3871 |
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Description: Minimum order quantity: 1 unit of 50x1mL |
) HiScript II Q Select RT SuperMix for qPCR (+g DNA wiper) |
|
R233-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 244 |
) Ultra SYBR Green qPCR Master Mix (2X, with ROX I) |
|
W2601-1 |
101Bio |
- |
Ask for price |
Ultra SYBR Green qPCR Master Mix (2X, with ROX I) |
|
W2601-5 |
101Bio |
- |
Ask for price |
The automation reduces the chance of handbook liquid dealing with errors, making the MRD disk a gorgeous resolution to guarantee reproducibility in moderate-throughput, longitudinal gene quantification purposes.
