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.
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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 Top Green qPCR SuperMix |
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20-abx098033 |
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 Tip Green qPCR SuperMix |
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EUR 276.00
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EUR 1596.00
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 Library Quantification qPCR SuperMix |
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20-abx098896 |
Abbexa |
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 QCell-Eva One-Step qRT-PCR SuperMix Kit |
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K5054200 |
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200 reactions |
EUR 489.6 |
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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-Eva One-Step qRT-PCR SuperMix Kit |
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K5054400 |
Biochain |
400 reactions |
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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-Eva One-Step qRT-PCR SuperMix Kit |
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K5054004 |
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400 rxn |
EUR 635 |
 Pro QPCR SuperMix Kit - ROX premixed |
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K5056200 |
Biochain |
200 reactions |
EUR 225.6 |
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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. |
Pro QPCR SuperMix Kit - ROX premixed |
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K5056400 |
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400 reactions |
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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. |
 cDNA Synthesis SuperMix for qPCR |
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20-abx09801920ulSystems |
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- 50 rxns × 20 ul Systems
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 Fast QCell-Eva One-Step qRT-PCR SuperMix Kit |
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K5054002 |
Biochain |
200 rxn |
EUR 418 |
 Fast Pro QPCR SuperMix Kit - ROX premixed |
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K5058200 |
Biochain |
200 reactions |
EUR 253.2 |
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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. |
Fast Pro QPCR SuperMix Kit - ROX premixed |
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K5058400 |
Biochain |
400 reactions |
EUR 384 |
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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. |
) Top Green qPCR SuperMix (+Dye II) |
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20-abx098890 |
Abbexa |
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EUR 994.80
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EUR 260.40
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EUR 1362.00
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EUR 493.20
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) Tip Green qPCR SuperMix (+Dye II) |
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20-abx098891 |
Abbexa |
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EUR 1161.60
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EUR 276.00
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) cDNA Synthesis SuperMix for qPCR (GC-rich) |
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abx098024-50rxns20ulSystems |
Abbexa |
50 rxns × 20 ul Systems |
EUR 861.6 |
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 HiScript II Q RT SuperMix for qPCR |
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R222-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 282 |
 HiScript II Q Select RT SuperMix for qPCR |
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R232-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 282 |
 PLATEMAX ULTRA CLEAR PERMANENT HEAT SEALING FILM FOR QPCR, 100/500 |
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HS-100-QPCR |
CORNING |
100/pk |
EUR 381.6 |
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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 381.6 |
|
Description: Sealing Products; Sealing films - Axygen |
) HiScript III RT SuperMix for qPCR (+g DNA wiper) |
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R323-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 351.6 |
 PCR SuperMix |
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20-abx098887 |
Abbexa |
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) HiScript II Q Select RT SuperMix for qPCR (+g DNA wiper) |
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R233-01 |
Vazyme |
100 rxn (20 μl/rxn) |
EUR 292.8 |
) PCR SuperMix (-dye) |
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20-abx098002 |
Abbexa |
-
EUR 493.20
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EUR 226.80
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EUR 309.60
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) PCR SuperMix (+dye) |
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20-abx098003 |
Abbexa |
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EUR 493.20
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EUR 226.80
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EUR 7408.80
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EUR 309.60
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 cDNA Synthesis SuperMix |
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20-abx09801420ulSystems |
Abbexa |
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- 100 rxns × 20 ul Systems
- 50 rxns × 20 ul Systems
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 Plus PCR SuperMix |
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20-abx098888 |
Abbexa |
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 Library Amplification SuperMix |
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20-abx098894 |
Abbexa |
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 Evo? cDNA Supermix |
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M1168-100 |
Biovision |
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EUR 457.2 |
Evo? cDNA Supermix |
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M1168-25 |
Biovision |
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EUR 320.4 |
 Novo? cDNA Supermix |
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M1169-100 |
Biovision |
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EUR 529.2 |
Novo? cDNA Supermix |
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M1169-25 |
Biovision |
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EUR 346.8 |
 OneScriptcDNA Synthesis SuperMix |
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G451 |
ABM |
25 x 20 ul reactions |
EUR 106.8 |
OneScriptcDNA Synthesis SuperMix |
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G452 |
ABM |
100 x 20 ul reactions |
EUR 169.2 |
) T PCR SuperMix (-dye) |
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20-abx098005 |
Abbexa |
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) T PCR SuperMix (+dye) |
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20-abx098006 |
Abbexa |
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) EasyPfu PCR SuperMix (-dye) |
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20-abx098009 |
Abbexa |
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) FastPfu PCR SuperMix (-dye) |
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20-abx098010 |
Abbexa |
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 OneScriptPlus cDNA Synthesis SuperMix |
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G453 |
ABM |
25 x 20 ul reactions |
EUR 116.4 |
OneScriptPlus cDNA Synthesis SuperMix |
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G454 |
ABM |
100 x 20 ul reactions |
EUR 202.8 |
 COLLECTION CONTAINER 1L, EVA FILM |
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91-200-01 |
CORNING |
1/pk |
EUR 96 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 2L, EVA FILM |
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91-200-02 |
CORNING |
1/pk |
EUR 105.6 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 5L, EVA FILM |
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91-200-05 |
CORNING |
1/pk |
EUR 118.8 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 10L, EVA FILM |
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91-200-10 |
CORNING |
1/pk |
EUR 121.2 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 20L, EVA FILM |
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91-200-20 |
CORNING |
1/pk |
EUR 132 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
COLLECTION CONTAINER 10L, EVA FILM |
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91-200-36 |
CORNING |
1/pk |
EUR 103.2 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
COLLECTION CONTAINER 20L, EVA FILM |
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91-200-39 |
CORNING |
1/pk |
EUR 106.8 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
COLLECTION CONTAINER 5L, EVA FILM |
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91-200-43 |
CORNING |
1/pk |
EUR 181.2 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
COLLECTION CONTAINER 10L, EVA FILM |
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91-200-45 |
CORNING |
1/pk |
EUR 151.2 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
COLLECTION CONTAINER 20L, EVA FILM |
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91-200-47 |
CORNING |
1/pk |
EUR 158.4 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 50L, EVA FILM |
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91-200-48 |
CORNING |
1/pk |
EUR 175.2 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 100L, EVA FILM |
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91-200-82 |
CORNING |
1/pk |
EUR 334.8 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 COLLECTION CONTAINER 200L, EVA FILM |
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91-200-83 |
CORNING |
1/pk |
EUR 350.4 |
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Description: Flexible Containers; Bioprocess Bags and Parts |
 Frit Kit |
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FRIT-KIT |
Next Advance |
1each |
EUR 148.8 |
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Description: Kit to create frits in capillaries. Includes formamide, Kasil-1, Kasil-1624 and a cleaving tool. |
 Column Packing Kit |
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PACK-KIT |
Next Advance |
1pack |
EUR 1242 |
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Description: Column packing kit for pressure cells. Includes: HPREG regulator, TBNG10 tubing, CAP-75 capillary, and STRB5X2 stir bar. |
 PCR Mycoplasma Detection Kit |
|
M034-Kit |
TOKU-E |
Kit |
EUR 319.2 |
 COLLECTION CONTAINER 500 ML, EVA FILM |
|
91-200-41 |
CORNING |
1/pk |
EUR 82.8 |
|
Description: Flexible Containers; Bioprocess Bags and Parts |
) PCR SuperMix for PAGE (+dye) |
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20-abx098004 |
Abbexa |
-
EUR 493.20
-
EUR 226.80
-
EUR 309.60
|
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 QPCR Positive Control Kit |
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M1126-100 |
Biovision |
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EUR 483.6 |
 Lentivirus qPCR Quantification Kit |
|
K1471-100 |
Biovision |
100 Rxns |
EUR 357.6 |
 Retrovirus qPCR Quantification Kit |
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K1472-100 |
Biovision |
100 Rxns |
EUR 381.6 |
 qPCR Control Template Kit |
|
G929 |
ABM |
100 rxns |
EUR 177.6 |
 Entrans qPCR Probe Kit |
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RK21209 |
Abclonal |
40 RXN |
Ask for price |
|
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 Cas9 Protein and T7 gRNA SmartNuclease Synthesis Kit |
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CAS400A-KIT |
SBI |
1 kit (10 rxn) |
EUR 1332 |
|
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 QCell-Pro One-Step qRT-PCR SuperMix Kit |
|
K5055200 |
Biochain |
200 reactions |
EUR 451.2 |
|
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 |
Biochain |
400 reactions |
EUR 686.4 |
|
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 |
|
K5055004 |
Biochain |
400 rxn |
EUR 609 |
 CMV-hspCas9-T2A-Puro SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit |
|
CASLV100PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
 CMV-hspCas9-EF1-GFP SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit |
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CASLV105PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
 MSCV-hspCas9-T2A-Puro SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit |
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CASLV120PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
 MSCV-hspCas9-EF1-GFP SmartNuclease Lentivector Plasmid + LentiStarter Packaging Kit |
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CASLV125PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
 Multiplex gRNA Kit + EF1-T7-hspCas9-H1-gRNA linearized SmartNuclease vector |
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CAS700A-KIT |
SBI |
10 rxn |
EUR 1358.4 |
|
|
 Multiplex gRNA Kit + CAG-T7-hspCas9-H1-gRNA linearized SmartNuclease vector |
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CAS720A-KIT |
SBI |
10 rxn |
EUR 1358.4 |
|
|
 Multiplex gRNA Kit + CMV-T7-hspCas9-H1-gRNA linearized SmartNuclease vector |
|
CAS740A-KIT |
SBI |
10 rxn |
EUR 1358.4 |
|
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) T7 gRNA SmartNuclease Synthesis Kit (includes CAS510A-1 & T7 IVT synthesis reagents) |
|
CAS510A-KIT |
SBI |
1 Kit |
EUR 966 |
|
|
-T2A-Puro SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit) Cas9 Nickase: CMV-hspCas9(D10A)-T2A-Puro SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit |
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CASLV200PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
-EF1-GFP SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit) Cas9 Nickase: CMV-hspCas9(D10A)-EF1-GFP SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit |
|
CASLV205PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
-T2A-Puro SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit) Cas9 Nickase: MSCV-hspCas9(D10A)-T2A-Puro SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit |
|
CASLV220PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
-EF1-GFP SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit) Cas9 Nickase: MSCV-hspCas9(D10A)-EF1-GFP SmartNickase Lentivector Plasmid + LentiStarter Packaging Kit |
|
CASLV225PA-KIT |
SBI |
1 Kit |
EUR 1358.4 |
|
|
 Multiplex gRNA Kit + Cas9 Nickase: EF1-T7-hspCas9-nickase-H1-gRNA linearized SmartNickase vector |
|
CAS750A-KIT |
SBI |
10 rxn |
EUR 1358.4 |
|
|
 Multiplex gRNA Kit + Cas9 Nickase: CAG-T7-hspCas9-nickase-H1-gRNA linearized SmartNickase vector |
|
CAS770A-KIT |
SBI |
10 rxn |
EUR 1358.4 |
|
|
 Multiplex gRNA Kit + Cas9 Nickase: CMV-T7-hspCas9-nickase-H1-gRNA linearized SmartNickase vector |
|
CAS790A-KIT |
SBI |
10 rxn |
EUR 1358.4 |
|
|
 Antibody) Protein Eva-1 Homolog C (EVA1C) Antibody |
|
20-abx302696 |
Abbexa |
-
EUR 493.20
-
EUR 2214.00
-
EUR 718.80
-
EUR 218.40
-
EUR 360.00
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- 100 ug
- 1 mg
- 200 ug
- 20 ug
- 50 ug
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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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