Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies

Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies

Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies

Microfluidic platforms provide a drastic enhance in throughput whereas minimizing pattern utilization and hands-on time, which make them essential instruments for large-scale organic research. A spread of such methods have been developed for enzyme exercise research, though their complexity largely hinders their software to the broader scientific group.

Right here, we current adaptation of an easy-to-use business microfluidic qPCR system for performing enzyme kinetic research. We display the performance of the Fluidigm Biomark HD system (the Fluidigm system) by figuring out the kinetic properties of three oxidases in a resorufin-based fluorescence assay. The outcomes obtained within the microfluidic system proved reproducible and akin to those obtained in a normal microplate-based assay. With a variety of easy-to-use, off-the-shelf elements, the microfluidic system presents itself as a easy and customizable platform for high-throughput enzyme exercise research.

A novel TaqMan qPCR assay for speedy detection and quantification of pro-inflammatory microalgae Prototheca spp. in milk samples

Animal or human protothecosis belongs to relatively uncommon, endemic, pro-inflammatory infections. It’s brought on by achlorophyllous algae of the genus Prototheca. Particularly, P. bovis (previously P. zopfii genotype 2) is usually inflected as a non-bacterial causative agent of dairy cattle mastitis. On this research, we current a multiplex real-time PCR (qPCR) system for speedy and precise Prototheca spp. detection and quantification. Restrict of detection, diagnostic sensitivity, and specificity have been decided.
For the primary time, particular sequences of AccD (encoding acetyl CoA reductase) for P. bovis, cox1 (encoding cytochrome C oxidase subunit 1) for P. wickerhamii, cytB (encoding cytochrome B) for P. blashkeae and atp6 (encoding transporting ATPase F0 subunit 6) for P. ciferrii (previously P. zopfii genotype 1) have been used for species identification and quantification along with 28S rRNA sequence detecting genus Prototheca. The developed qPCR assay was utilized to 55 particular person cow milk samples from a herd suspected of protothecosis, 41 bulk milk samples from completely different Czech farms, 16 boxed milk samples bought in supermarkets and 21 environmental samples originating from a farm suspected of protothecosis. Our work thus presents the chance to diagnose protothecosis within the samples, the place bacterial mastitis is essentially the most generally presumed and thereby helping ample corrective measures to be taken.

Lowered ribosomal DNA transcription within the prefrontal cortex of suicide victims: consistence of latest molecular RT-qPCR findings with earlier morphometric information from AgNOR-stained pyramidal neurons

Prefrontal cortical areas play a key function in behavioural regulation, which is profoundly disturbed in suicide. The research was carried out on frozen cortical samples from the anterior cingulate cortex (dorsal and ventral elements, ACd and ACv), the orbitofrontal cortex (OFC), and the dorsolateral cortex (DLC) obtained from 20 suicide completers (predominantly violent) with unknown psychiatric analysis and 21 non-suicidal controls. The relative degree of ribosomal RNA (rRNA) as a marker of the transcriptional exercise of ribosomal DNA (rDNA) was evaluated bilaterally in prefrontal areas talked about above (i.e. in eight areas of curiosity, ROIs) by reverse transcription and quantitative polymerase chain response (RT-qPCR).
The general statistical evaluation revealed a lower in rDNA exercise in suicide victims versus controls, notably in male topics. Additional ROI-specific submit hoc analyses revealed a major lower on this exercise in suicides in comparison with non-suicides in 5 ROIs. This impact was accentuated within the ACv, the place it was noticed bilaterally. Our findings counsel that decreased rDNA transcription within the prefrontal cortex performs an essential function in suicide pathogenesis and corresponds with our earlier morphometric analyses of AgNOR-stained neurons.
Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies

Choice and validation of inside reference genes for qPCR in Polygonatum cyrtonema tubers at completely different growth levels and in response to abiotic stress

With a purpose to analyze the expression of genes concerned in steroidal saponin biosynthesis pathway in Polygonatum cyrtonema tubers, it is rather essential to pick out inside reference genes which might be stably expressed at completely different growth levels and in response to abiotic stress. Based on the beforehand established P. cyrtonema transcriptome database and reported inside reference genes in plant, this research systematically analyzed eight candidate inside reference genes together with histone H2 A, glyceraldehyde-3-phosphate dehydrogenase, ACTIN, β-tubulin, ubiquitin-conjugating enzyme-E2-10, elongation issue 1-alpha isoform, 18 S rRNA and α-tubulin four for expression stability in P. cyrtonema tubers at completely different growth levels and in response to methyl jasmonate(MeJA) stress by utilizing Actual time fluorescence quantitative PCR(qPCR). Based mostly on the statistical evaluation of qPCR outcomes by utilizing GeNorm, NormFinder and BestKeeper softwares, the expression of ubiquitin-conjugating enzyme-E2-10 and elongation issue 1-alpha isoform are essentially the most steady in P. cyrtonema tubes at completely different growth levels and in response to MeJA stress.

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HotTaq Probe qPCR Mix (ROX)

BT11001 250rxn
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HotTaq Probe qPCR Mix (Capillary)

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HotTaq Probe qPCR Universal Mix

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HotTaq EvaGreen qPCR Mix (ROX)

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HotTaq EvaGreen qPCR Mix (Capillary)

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HotTaq EvaGreen qPCR Universal Mix

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SYBR Green qPCR Master Mix

HY-K0501 5 mL (500 rxns)
EUR 263

RT Master Mix for qPCR

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EUR 291

BrightGreen 5X qPCR MasterMix-iCycler

MasterMix-5C 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

TaqProbe 5X qPCR MasterMix-ROX

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

TaqProbe 5X qPCR MasterMix-iCycler

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EUR 213

TaqProbe 5X qPCR MasterMix-Multiplex

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EUR 213

BrightGreen 5X qPCR MasterMix-ROX

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

BrightGreen 2X qPCR MasterMix-iCycler

MasterMix-iC 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-iCycler

MasterMix-iC-XL 16 x 1.25 ml for 2000 reactions (20 ul)
EUR 376

KiloGreen 2X qPCR MasterMix-iCycler

MasterMix-KC 4 x 1.25 ml - 500 reactions (20 ul)
EUR 140

KiloGreen 2X qPCR MasterMix-ROX

MasterMix-KR 4 x 1.25 ml - 500 reactions (20 ul)
EUR 140

BrightGreen miRNA qPCR MasterMix-iCycler

MasterMix-mC 4 x 1.25 ml for 500 reactions
EUR 151

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

TaqProbe 2X qPCR MasterMix-iCycler

MasterMix-PC 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

TaqProbe 2X qPCR MasterMix-Multiplex

MasterMix-PM 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

AceQ qPCR Probe Master Mix

Q112-02 500 rxn (20 μl/rxn)
EUR 221

AceQ qPCR Probe Master Mix

Q112-03 2500 rxn (20 μl/rxn)
EUR 646

AAVS1 Safe Harbor Targeting Vector 2.0 - All-Purpose Donor (AAVS1-SA-puro-MCS), Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector) and GE640PR-1 (Junction PCR Primer Mix to confirm AAVS1 integration site)

GE620A-KIT 1 kit
EUR 2132
  • Category: Gene Editing

AAVS1 Safe Harbor Targeting Vector 2.0 - GOI Knock-in Donor (AAVS1-SA-puro-EF1-MCS), Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector) and GE640PR-1 (Junction PCR Primer Mix to confirm AAVS1 integration site)

GE622A-KIT 1 kit
EUR 2132
  • Category: Gene Editing

AAVS1 Safe Harbor Targeting Vector 2.0 - Reporter Knock-in Donor (AAVS1-SA-puro-MCS-GFP), Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector) and GE640PR-1 (Junction PCR Primer Mix to confirm AAVS1 integration site)

GE624A-KIT 1 kit
EUR 2132
  • Category: Gene Editing

First-Strand cDNA Synthesis SuperMix for PCR

abx098018-50rxns20ulSystems 50 rxns × 20 ul Systems
EUR 578
  • Shipped within 5-10 working days.

Two-Step RT-PCR SuperMix (12 kb)

abx098025-50rxns20ulRTSystems80rxns50ulPCRSystems 50 rxns × 20 ul (RT Systems) / 80 rxns × 50 ul (PCR Systems)
EUR 606
  • Shipped within 5-10 working days.

Two-Step RT-PCR SuperMix (15 kb)

abx098026-50rxns20ulRTSystems80rxns50ulPCRSystems 50 rxns × 20 ul (RT Systems) / 80 rxns × 50 ul (PCR Systems)
EUR 759
  • Shipped within 5-10 working days.

Green miRNA Two-Step qRT-PCR SuperMix

abx098036-20rxns20ulRTSystems500rxns20ulqPCRSystems 20 rxns × 20 ul (RTSystems) / 500 rxns × 20 ul (qPCRSystems)
EUR 885
  • Shipped within 5-10 working days.

igScript One Step RT-qPCR Kit - 100 Reactions

4214 1/EA
EUR 253

igScript One Step RT-qPCR Kit - 500 Reactions

4218 1/EA
EUR 773

One.Step RT-qPCR Kit with EvaGreen (2X concentrated)

105-520 2 x 1,25 ml
EUR 458

One.Step RT-qPCR Kit with EvaGreen (2X concentrated)

105-522 10 x 1,25 ml
EUR 1719

vWF Acty. Kit

ABP-ACT-KIT 12 x 8 microwells
EUR 428

vWF Ant. Kit

ABP-TOT-KIT 12 x 8 microwells
EUR 394

hspCas9 AAVS1 Safe Harbor Knock-in Donor (AAVS1-SA-puro-EF1-hspCas9)

CAS620A-KIT 1 kit
EUR 2152
  • Category: Cas9
Description: Complete Kit with CAS601A-1 (Cas9 SmartNuclease AAVS1-gRNA Targeting Vector), CAS640PR-1 (Junction PCR Primer Mix to confirm Cas9 integration site), and CAS9-PR-1 (PCR primers to confirm Cas9 expression)

PinPoint-FC System for Platform Cell Line Generation & Retargeting (includes PIN300A-1, FC200PA-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN300A-KIT 1 Kit
EUR 2798
  • Category: PinPoint Integrase Tools

PinPoint-HR System for Platform Cell Line Generation & Retargeting (includes PIN400A-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN400A-KIT 1 Kit
EUR 2798
  • Category: PinPoint Integrase Tools

PinPoint-HR System for Platform Cell Line Generation & Retargeting of AAVS1 Safe Harbor Locus (includes PIN410A-1, GE601A-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN410A-KIT 1 Kit
EUR 4335
  • Category: PinPoint Integrase Tools

PinPoint-HR System for Platform Cell Line Generation & Retargeting of AAVS1 Safe Harbor Locus (includes PIN410A-1, CAS601A-1, PIN200A-1, PIN510A-1, & PIN600A-1)

PIN412A-KIT 1 Kit
EUR 4335
  • Category: PinPoint Integrase Tools

2x SYBR Green qPCR Master Mix

B21202 5 mL
EUR 224
Description: Our 2x SYBR Green qPCR master mix performs toe to toe in all qPCR assays with the most well known brands on the market but surpases them significantly in cost-efficiency.

2x SYBR Green qPCR Master Mix

B21203 25 mL
EUR 856
Description: Our 2x SYBR Green qPCR master mix performs toe to toe in all qPCR assays with the most well known brands on the market but surpases them significantly in cost-efficiency.

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.

Opti-Seal qPCR and Storage Films

G495 100 Films/Box
EUR 202

Opti-Seal qPCR 384 Well Plate

G935 10 Plates/Box
EUR 89

Opti-Seal qPCR 384 Well Plate

G936 100 Plates/Case
EUR 428

Universal miRNA qPCR Primer (10 µM)

abx298006-200ul 200 ul
EUR 189
  • Shipped within 5-10 working days.

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

TaqProbe 5X qPCR MasterMix-No Dye

MasterMix-5PS 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

BrightGreen 5X qPCR MasterMix-No Dye

MasterMix-5S 4 x 1.0 ml for 1000 reactions (20 ul)
EUR 213

BrightGreen Express 2X qPCR MasterMix-iCycler

MasterMix-EC 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

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

KiloGreen 2X qPCR MasterMix-No Dye

MasterMix-KS 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

BrightGreen miRNA qPCR MasterMix-No Dye

MasterMix-mS 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

TaqProbe 2X qPCR MasterMix-No Dye

MasterMix-PS 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-No Dye

MasterMix-S 4 x 1.25 ml for 500 reactions (20 ul)
EUR 140

BrightGreen 2X qPCR MasterMix-No Dye

MasterMix-S-XL 16 x 1.25 ml for 2000 reactions (20 ul)
EUR 376

AceQ Universal SYBR qPCR Master Mix

Q511-02 500 rxn (20 μl/rxn)
EUR 221

AceQ Universal SYBR qPCR Master Mix

Q511-03 2500 rxn (20 μl/rxn)
EUR 646

ChamQ Universal SYBR qPCR Master Mix

Q711-02 500 rxn (20 μl/rxn)
EUR 221

ChamQ Universal SYBR qPCR Master Mix

Q711-03 2500 rxn (20 μl/rxn)
EUR 646

PrecisionX Multiplex gRNA Cloning Kit

CAS9-GRNA-KIT 10 rxn
EUR 445
  • Category: Cas9

High Fidelity (HiFi) PCR SuperMix (-dye) (DNA, PCR)

20-abx098007
  • EUR 217.00
  • EUR 356.00
  • 1 ml
  • 5 ml
  • Shipped within 5-10 working days.

One-Step RT-PCR SuperMix (+dye) (4 kb)

abx098027-200rxns20ulSystems 200 rxns × 20 ul Systems
EUR 606
  • Shipped within 5-10 working days.

One-Step RT-PCR SuperMix (+dye) (8 kb)

abx098028-200rxns20ulSystems 200 rxns × 20 ul Systems
EUR 662
  • Shipped within 5-10 working days.

One-Step RT-PCR SuperMix (+dye) (8 kb)

abx098029-200rxns20ulSystems 200 rxns × 20 ul Systems
EUR 857
  • Shipped within 5-10 working days.

Green Two-Step qRT-PCR SuperMix (GC Rich)

abx098037-50rxns20ulRTSystems300rxns20ulqPCRSystems 50 rxns × 20 ul (RTSystems) / 300 rxns × 20 ul (qPCRSystems)
EUR 801
  • Shipped within 5-10 working days.

Green One-Step qRT-PCR SuperMix (GC Rich)

20-abx09803920ulSystems
  • EUR 592.00
  • EUR 1024.00
  • 100 rxns × 20 ul Systems
  • 400 rxns × 20 ul Systems
  • Shipped within 5-10 working days.

Probe One-Step qRT-PCR SuperMix (GC Rich)

20-abx09804120ulSystems
  • EUR 495.00
  • EUR 746.00
  • 100 rxns × 20 ul Systems
  • 400 rxns × 20 ul Systems
  • Shipped within 5-10 working days.

One-Step gDNA Removal and cDNA Synthesis SuperMix

20-abx09885920ulSystems
  • EUR 1024.00
  • EUR 746.00
  • 100 rxns × 20 ul Systems
  • 50 rxns × 20 ul Systems
  • Shipped within 5-10 working days.

One.Step RT-qPCR Probe Kit with HighROX (2X concentrated)

105-510 2 x 1,25 ml
EUR 440

One.Step RT-qPCR Probe Kit with HighROX (2X concentrated)

105-512 10 x 1,25 ml
EUR 1628

mRNAExpress mRNA Synthesis kit (5 reactions)

MR-KIT-1 5 reactions
EUR 1152
  • Category: Stem Cell Products

One-Step gDNA Removal and cDNA Synthesis SuperMix (15kb)

20-abx09802220ulSystems
  • EUR 940.00
  • EUR 690.00
  • 100 rxns × 20 ul Systems
  • 50 rxns × 20 ul Systems
  • Shipped within 5-10 working days.

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

Opti-Seal qPCR Semi-skirted plate (Standard)

G908 10 Plates/Bag
EUR 76

Opti-Seal qPCR Semi-skirted plate (Standard)

G911 100 Plates/Case
EUR 302

Opti-Seal qPCR Roche 480 (Low profile)

G913 100 Plates/Case
EUR 302

Opti-Seal qPCR Roche 480 (Low profile)

G914 10 Plates/Box
EUR 76
The 2 inside reference genes have been additional validated by analyzing the expression of four genes concerned in steroidal saponin biosynthesis pathways. In conclusion, ubiquitin-conjugating enzyme-E2-10 and elongation issue 1-alpha isoform can be utilized as essentially the most acceptable inside reference genes for qPCR evaluation in P. cyrtonema. This research additionally present a basis for future examine the molecular mechanism of steroidal saponin biosynthesis pathways in P. cyrtonema.

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