Enhanced asymmetric blocked qPCR method for affordable detection of point mutations in KRAS oncogene

Enhanced asymmetric blocked qPCR method for affordable detection of point mutations in KRAS oncogene

Enhanced asymmetric blocked qPCR method for affordable detection of point mutations in KRAS oncogene

An correct genetic diagnostic is essential for enough affected person administration and the suitability of healthcare techniques. The scientific problem lies in growing strategies to discriminate these sufferers with sure genetic variations current in tumor cells at low concentrations. We report a technique referred to as enhanced uneven blocked qPCR (EAB-qPCR) that promotes the blocker annealing towards the primer-template hybrid controlling thermal biking and response circumstances with nonmodified oligonucleotides. Actual-time fluorescent amplification curves of wild-type alleles had been delayed by about eight cycles for EAB-qPCR, in comparison with standard blocked qPCR approaches.

This methodology diminished the amplification of native DNA variants (blocking proportion 99.7%) and enabled the efficient enrichment of low-level DNA mutations. Wonderful efficiency was estimated for the detection of mutated alleles in sensitivity (as much as 0.5% mutant/whole DNA) and reproducibility phrases, with a relative commonplace deviation under 2.8%. The strategy was efficiently utilized to the mutational evaluation of metastatic colorectal carcinoma from biopsied tissues.

The decided single-nucleotide mutations within the KRAS oncogene (codon 12-13) completely agreed with these obtained from next-generation sequencing. EAB-qPCR is an correct low cost methodology and might be simply included into day by day routine to detect mutant alleles. Therefore, these options are particularly attention-grabbing to facilitate the analysis and prognosis of a number of scientific ailments.

Quantification of Actaea racemosa L. (black cohosh) from a few of its potential adulterants utilizing qPCR and dPCR strategies

The demand for well-liked pure well being merchandise (NHPs) similar to Black Cohosh is growing significantly, which in flip challenges high quality assurance (QA) all through the availability chain. To detect and quantify the goal species current in a given NHP, DNA-based molecular strategies similar to Actual-time quantitative PCR (qPCR) and digital PCR (dPCR) are commonplace instruments within the meals and pathogen testing industries.
There’s a hole within the literature regarding validated quantitative PCR strategies for botanicals that may be utilized for QA and good manufacturing practices. The target of this research is to develop an environment friendly quantification methodology utilizing qPCR and dPCR strategies for the detection and quantification of Actaea racemosa (Black cohosh) NHPs from its potential adulterants. These developed strategies are validated for applicability on business NHPs.
Species-specific hydrolysis probe assays had been designed to research the black cohosh NHPs utilizing qPCR and dPCR strategies. The outcomes confirmed that the developed qPCR and dPCR strategies are extremely exact for figuring out and quantifying black cohosh NHPs, indicating their potential applicability in future routine industrial and laboratory testing. This permits a single qPCR check to find out not solely the presence of a particular botanical, but additionally the quantity when combined with an adulterant.

Quantification of Legionella pneumophila by qPCR and tradition in faucet water with completely different concentrations of residual disinfectants and heterotrophic micro organism

Legionellosis prevalence is growing in the USA. This illness is precipitated primarily by the bacterium Legionella pneumophila present in water and transmitted by aerosol inhalation. This pathogen has a gradual progress fee and may “disguise” in amoeba, making it tough to watch by the normal tradition methodology on selective media. Faucet water samples (n = 358) collected throughout the USA had been examined for L. pneumophila by each tradition and quantitative Polymerase Chain Response (qPCR).
The presence of different micro organism was quantified by heterotrophic plate counts (HPC). Residual disinfectant concentrations (free chlorine or monochloramine) had been measured in all samples. Legionella pneumophila had the best prevalence and focus within the chlorinated water samples that had a free‑chlorine worth of lower than 0.2 mg Cl2/L. In whole, 24% (87/358) of the samples had been constructive for L. pneumophila both by qPCR or 3% (11/358) had been constructive by tradition.
Enhanced asymmetric blocked qPCR method for affordable detection of point mutations in KRAS oncogene
In chloramine-treated samples, L. pneumophila was detected by qPCR in 21% (31/148) and 1% (2/148) by tradition, regardless of a excessive monochloramine residual >1 mg Cl2/L. Regardless of the presence of a excessive disinfectant residual (>1 mg Cl2/L), HPC counts had been substantial. This research signifies that each tradition and qPCR strategies have limitations when predicting a possible danger for illness related to L. pneumophila in faucet water. Measuring disinfectant residuals and quantifying HPC in water samples could also be helpful adjunct parameters for lowering Legionellosis’ danger from public water provides at high-risk areas.

PANDAA deliberately violates standard qPCR design to allow sturdy, mismatch-agnostic detection of extremely polymorphic pathogens

Delicate and reproducible diagnostics are elementary to containing the unfold of present and rising pathogens. Regardless of the reliance of scientific virology on qPCR, technical challenges persist that compromise their reliability for sustainable epidemic containment as sequence instability in probe-binding areas produces false-negative outcomes.
We systematically violated canonical qPCR design rules to develop a Pan-Degenerate Amplification and Adaptation (PANDAA), a degree mutation assay that mitigates the influence of sequence variation on probe-based qPCR efficiency. Utilizing HIV-1 as a mannequin system, we optimized and validated PANDAA to detect HIV drug resistance mutations (DRMs). Extremely-degenerate primers with 3′ termini overlapping the probe-binding web site adapt the goal by site-directed mutagenesis throughout qPCR to exchange DRM-proximal sequence variation.

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Description: SYBR Green qPCR Master Mix (No ROX)

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Description: SYBR Green qPCR Master Mix (No ROX)

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Description: SYBR Green qPCR Master Mix (No ROX)

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Description: SYBR Green qPCR Master Mix (No ROX)

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Description: SYBR Green qPCR Master Mix (No ROX)

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Description: SYBR Green qPCR Master Mix (Low ROX)

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Description: SYBR Green qPCR Master Mix (Low ROX)

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Description: SYBR Green qPCR Master Mix (Low ROX)

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Description: SYBR Green qPCR Master Mix (Low ROX)

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Description: SYBR Green qPCR Master Mix (Low ROX)

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HY-K0522 5 mL (500 rxns )
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Description: SYBR Green qPCR Master Mix (High ROX)

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Description: SYBR Green qPCR Master Mix (High ROX)

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Description: SYBR Green qPCR Master Mix (High ROX)

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Description: SYBR Green qPCR Master Mix (High ROX)

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Description: SYBR Green qPCR Master Mix (High ROX)

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EUR 135.6

2x SYBR Green qPCR Master Mix

B21202 5 mL
EUR 268.8
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ig SYBR Green qPCR 2X Master Mix - 200 Reactions

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Description: qPCR Kit, Universal SYBR Green I Master Mix (Rox)

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17270 1 mL
EUR 203
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EUR 508
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17274 1 mL
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TAQuestâ„¢ qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

17274-1mL 1 mL
EUR 99
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TAQuestâ„¢ qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

17275 5 mL
EUR 508
Description: TAQuestâ„¢ qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

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17276 1 mL
EUR 203
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TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *No ROX*

17276-1mL 1 mL
EUR 99
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *No ROX*

17277 5 mL
EUR 508
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

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17277-5mL 5 mL
EUR 308
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *Low ROX*

17278 1 mL
EUR 203
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

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17278-1mL 1 mL
EUR 99
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *Low ROX*

17279 5 mL
EUR 508
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

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17279-5mL 5 mL
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Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

17280 1 mL
EUR 203
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

17280-1mL 1 mL
EUR 99
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

17281 5 mL
EUR 508
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green *High ROX*

17281-5mL 5 mL
EUR 308
Description: TAQuestâ„¢ FAST qPCR Master Mix with Helixyteâ„¢ Green is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR.

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Q331-02 500 rxns (20 μl/rxn)
EUR 120.55

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EUR 543.5

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EUR 120.55

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Q341-03 2,500 rxns (20 μl/rxn)
EUR 543.5

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RK21205 1 ml
EUR 328.5

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Q431-02 500 rxns (20 μl/rxn)
EUR 140.63

ChamQ SYBR Color qPCR Master Mix (Low ROX Premixed)

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EUR 634

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Q441-02 500 rxns (20 μl/rxn)
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EUR 328.5

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EUR 120.55

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Q311-03 2,500 rxns (20 μl/rxn)
EUR 543.5

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Z-PPLUS-R-1ML 1ml
EUR 81
Description: Precision®PLUS

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Apex qPCR GREEN Master Mix

42-116PG 400 x 25µl Reactions/Unit
EUR 341.44
Description: 2X, Without ROX, 5ml

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42-117PG 4000 x 25µl Reactions/Unit
EUR 2926.74
Description: 2X, Without ROX, 50ml

Apex qPCR GREEN Master Mix

42-118PG 400 x 25µl Reactions/Unit
EUR 338.88
Description: 2X, Low ROX, 5ml

Apex qPCR GREEN Master Mix

42-119PG 4000 x 25µl Reactions/Unit
EUR 2926.74
Description: 2X, Low ROX, 50ml

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42-120PG 400 x 25µl Reactions/Unit
EUR 341.44
Description: 2X, High ROX, 5ml

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42-121PG 4000 x 25µl Reactions/Unit
EUR 3095.57
Description: 2X, High ROX, 50ml

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1ml Precision FAST qPCR Master Mix with ROX

Z-PFAST-R-1ML 1ml
EUR 90
Description: Precision®FAST

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Z-PFAST-R-2ML 2ml
EUR 161
Description: Precision®FAST

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EUR 140.63

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EUR 152.72

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EUR 688.5

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RK21204 1 mL
EUR 328.5

iGreen 2X qPCR Master Mix with Low-ROX, 500Rxns-20ul

A4183-4x125ML500Rxns 4x1.25ML (500Rxns)
EUR 200.2
Description: Biotechnology

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RK21203 1 mL
EUR 76.5

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RK21219 5mL
EUR 475

Hi-SYBr Master Mix (with ROX)

MBT160-0.5ML 1 unit
EUR 52.29
Description: Hi-SYBr Master Mix (with ROX)

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EUR 94.08
Description: Hi-SYBr Master Mix (with ROX)

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EUR 376.4
Description: Hi-SYBr Master Mix (with ROX)

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lyophilised real-time PCR Master Mix with SYBR® Green fluorescent DNA stain

M-PCR-173L 960 reactions x 20 µl
EUR 897
Description: lyophilised real-time PCR Master Mix with SYBR® Green fluorescent DNA stain

lyophilised real-time PCR Master Mix with SYBR® Green fluorescent DNA stain

M-PCR-173S 192 reactions x 20 µl
EUR 224
Description: lyophilised real-time PCR Master Mix with SYBR® Green fluorescent DNA stain

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EUR 161

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Q712-03 2,500 rxns (20 μl/rxn)
EUR 725

2ml PrecisionPLUS qPCR Master Mix with ROX (2 x 1ml)

Z-PPLUS-R-2ML 2ml
EUR 140
Description: Precision®PLUS

5ml PrecisionPLUS qPCR Master Mix with ROX (5 x 1ml)

Z-PPLUS-R-5ML 5ml
EUR 296
Description: Precision®PLUS

10ml PrecisionPLUS qPCR Master Mix with ROX (10 x 1ml)

Z-PPLUS-R-10ML 10ml
EUR 530
Description: Precision®PLUS

20ml PrecisionPLUS qPCR Master Mix with ROX (20 x 1ml)

Z-PPLUS-R-20ML 20ml
EUR 1025
Description: Precision®PLUS

5ml Precision FAST qPCR Master Mix with ROX (5 x 1ml)

Z-PFAST-R-5ML 5ml
EUR 348
Description: Precision®FAST

10ml Precision FAST qPCR Master Mix with ROX (10 x 1ml)

Z-PFAST-R-10ML 10ml
EUR 641
Description: Precision®FAST

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Z-PFAST-R-20ML 20ml
EUR 1120
Description: Precision®FAST

Entrans 2X qPCR Probe Master Mix with UDG

RK21222 50RXN
EUR 71

Fast Plus EvaGreen® qPCR Master Mix with low Rox (5000 rxn)

31014-2 50x1mL
EUR 3970
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Fast Plus EvaGreen® qPCR Master Mix with low Rox (5000 rxn)

31014-2-1 PK
EUR 3970

Fast Plus EvaGreen® qPCR Master Mix with high Rox (5000 rxn)

31015-2 50x1mL
EUR 3970
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Fast Plus EvaGreen® qPCR Master Mix with high Rox (5000 rxn)

31015-2-1 PK
EUR 3970

ViPrimePLUS Taq qPCR Green Master Mix II, 150rxns

QLMM17 each Ask for price

Fast Plus EvaGreen® qPCR Master Mix with low Rox (500 rxn)

31014-1-1 PK
EUR 437

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31015-1-1 PK
EUR 437

ViPrimePLUS AtTaq qPCR Green Master Mix II, 150rxns

QLMM07 each Ask for price

1ml PrecisionPLUS qPCR Master Mix premixed with SYBRgreen

Z-PPLUS-SY-1ML 1ml
EUR 81
Description: Precision®PLUS

1ml Precision FAST qPCR Master Mix premixed with SYBRgreen

Z-PFAST-SY-1ML 1ml
EUR 90
Description: Precision®FAST

2ml Precision FAST qPCR Master Mix premixed with SYBRgreen

Z-PFAST-SY-2ML 2ml
EUR 161
Description: Precision®FAST

Forget-Me-Not™ EvaGreen® qPCR Master Mix with ROX (2000 reactions)

31042-20mL 2x10mL
EUR 500
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Forget-Me-Not™ EvaGreen® qPCR Master Mix with ROX (2000 reactions)

31042-20mL-1 PK
EUR 500

Forget-Me-Not™ EvaGreen® qPCR Master Mix with ROX (500 reactions)

31042-1 5x1mL
EUR 251
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Forget-Me-Not™ EvaGreen® qPCR Master Mix with ROX (500 reactions)

31042-1-1 PK
EUR 251

Forget-Me-Not™ EvaGreen® qPCR Master Mix with ROX (100 reactions)

31042-T 1mL
EUR 73
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Forget-Me-Not™ EvaGreen® qPCR Master Mix with ROX (100 reactions)

31042-T-1 EA
EUR 73

Maximo Dry-Master Mix (Lyophilized qPCR Mastermix with Sybrgreen)

S295RS 192 rcs (20µl, in plates or tubes)
EUR 171

amfiSure Ultra Fidelity PCR Master Mix(2X)

P0346-001 1ml
EUR 336

amfiSure Ultra Fidelity PCR Master Mix(2X)

P0346-002 2x1ml
EUR 578.4

amfiSure Ultra Fidelity PCR Master Mix(2X)

P0346-005 5x1ml
EUR 993.6

amfiSure Ultra Fidelity PCR Master Mix(2X)

P0346-010 10x1ml
EUR 1632

Fast Plus EvaGreen® qPCR Master Mix with low Rox (trial size, 100 rxn)

31014-T 1mL
EUR 94
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Fast Plus EvaGreen® qPCR Master Mix with low Rox (trial size, 100 rxn)

31014-T-1 EA
EUR 94

Fast Plus EvaGreen® qPCR Master Mix with High Rox (trial size, 100 rxn)

31015-T 1mL
EUR 94
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Fast Plus EvaGreen® qPCR Master Mix with High Rox (trial size, 100 rxn)

31015-T-1 EA
EUR 94

Fast Plus EvaGreen® Master Mix for qPCR with low Rox (200 rxn)

31014-1 5x1mL
EUR 201
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

Fast Plus EvaGreen® Master Mix for qPCR with high Rox (200 rxn)

31015-1 5x1mL
EUR 201
Description: BSA from bovine serum (cow; Bos taurus); recombinant Taq polymerase from E. coli

HotStart PCR Master mix (2X, Green Dye)

BS9291 5X1mL, 5ml
EUR 251.4

HotStart PCR Master mix (2X, Green Dye)

BS9292 1ml
EUR 115.68

amfiSure qGreen Q-PCR Master Mix(2X), No Rox

Q5600-001 1ml
EUR 175.2

amfiSure qGreen Q-PCR Master Mix(2X), No Rox

Q5600-002 2x1ml
EUR 248.4

amfiSure qGreen Q-PCR Master Mix(2X), No Rox

Q5600-005 5x1ml
EUR 322.8

amfiSure qGreen Q-PCR Master Mix(2X), No Rox

Q5600-010 10x1ml
EUR 537.6

amfiSure qGreen Q-PCR Master Mix(2X), No Rox

Q5600-050 50x1ml
EUR 2085.6

amfiSure qProbe Q-PCR Master Mix(2X), No Rox

Q5700-001 1ml
EUR 208.8

amfiSure qProbe Q-PCR Master Mix(2X), No Rox

Q5700-002 2x1ml
EUR 319.2

amfiSure qProbe Q-PCR Master Mix(2X), No Rox

Q5700-005 5x1ml
EUR 562.8

amfiSure qProbe Q-PCR Master Mix(2X), No Rox

Q5700-010 10x1ml
EUR 886.8

amfiSure qProbe Q-PCR Master Mix(2X), No Rox

Q5700-050 50x1ml
EUR 3462

amfiSure qGreen Q-PCR Master Mix(2X), Low Rox

Q5603-001 1ml
EUR 175.2

amfiSure qGreen Q-PCR Master Mix(2X), Low Rox

Q5603-002 2x1ml
EUR 248.4

amfiSure qGreen Q-PCR Master Mix(2X), Low Rox

Q5603-005 5x1ml
EUR 319.2

amfiSure qGreen Q-PCR Master Mix(2X), Low Rox

Q5603-010 10x1ml
EUR 530.4

amfiSure qGreen Q-PCR Master Mix(2X), Low Rox

Q5603-050 50x1ml
EUR 2004

amfiSure qProbe Q-PCR Master Mix(2X), Low Rox

Q5703-001 1ml
EUR 208.8

amfiSure qProbe Q-PCR Master Mix(2X), Low Rox

Q5703-002 2x1ml
EUR 319.2

amfiSure qProbe Q-PCR Master Mix(2X), Low Rox

Q5703-003 5x1ml
EUR 562.8

amfiSure qProbe Q-PCR Master Mix(2X), Low Rox

Q5703-004 10x1ml
EUR 886.8

amfiSure qProbe Q-PCR Master Mix(2X), Low Rox

Q5703-005 50x1ml
EUR 3462

amfiSure qProbe Q-PCR Master Mix(2X), Low Rox

Q5703-050 50x1ml
EUR 1910

PanGreen™ Universal SYBR® Green Mastermix w Low ROX

QSD01-L100 100 rxns
EUR 20

Green-2-Go qPCR Mastermix-Low ROX (500X20ul Rxn)

QPCR002-L 4X1.25ml
EUR 303.25

amfiSure qGreen Q-PCR Master Mix(2X), High Rox

Q5602-001 1ml
EUR 175.2

amfiSure qGreen Q-PCR Master Mix(2X), High Rox

Q5602-002 2x1ml
EUR 248.4

amfiSure qGreen Q-PCR Master Mix(2X), High Rox

Q5602-005 5x1ml
EUR 319.2

amfiSure qGreen Q-PCR Master Mix(2X), High Rox

Q5602-010 10x1ml
EUR 530.4

amfiSure qGreen Q-PCR Master Mix(2X), High Rox

Q5602-050 50x1ml
EUR 2004

amfiSure qProbe Q-PCR Master Mix(2X), High Rox

Q5702-001 1ml
EUR 208.8

amfiSure qProbe Q-PCR Master Mix(2X), High Rox

Q5702-002 2x1ml
EUR 319.2

amfiSure qProbe Q-PCR Master Mix(2X), High Rox

Q5702-005 5x1ml
EUR 562.8

amfiSure qProbe Q-PCR Master Mix(2X), High Rox

Q5702-010 10x1ml
EUR 886.8

amfiSure qProbe Q-PCR Master Mix(2X), High Rox

Q5702-050 50x1ml
EUR 400.8

PanGreen™ Universal SYBR® Green Mastermix w High ROX

QSD01-H100 100 rxns
EUR 20

PanGreen™ Universal SYBR Green Mastermix UNG w Low ROX

QSD02-L100 100 rxns
EUR 22

1-Step 2X RT-PCR Master Mix-SYBR Green-HY (High Yield)

W141 200 rxn
EUR 199

1ml PrecisionPLUS OneStep RT-qPCR Master Mix with ROX

Z-OSPLUS-R-1ML 1ml
EUR 179
Description: Precision®PLUS OneStep

Entrans 2X qPCR Probe Master Mix

RK21208 40 RXN
EUR 119

PanGreen™ Universal SYBR Green Mastermix UNG w High ROX

QSD02-H100 100 rxns
EUR 22

Accuris qMax Green, qPCR mix with blue tracking dye, No Rox qPCR Mix, 1000 reactions

PR2000-N-1000 1 PC
EUR 642.51

ViPrimePLUS OneStep Taq RT-qPCR Master Mix with ROX, 150rxns

QLMM13-R each Ask for price
PANDAA-quantified DRMs current at frequency ≥5% (2 h from nucleic acid to consequence) with a sensitivity and specificity of 96.9% and 97.5%, respectively. PANDAA is an revolutionary development with applicability to any pathogen the place target-proximal genetic variability hinders diagnostic growth.

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