The elevated utilization of high-throughput applied sciences in most cancers analysis, together with genetic and drug screens, generates massive units of candidate targets that should be functionally validated for his or her roles in tumor growth. Thus, dependable and strong in vivo mannequin methods are wanted to carry out reverse genetic experiments. Ideally, these fashions ought to enable for a conditional silencing of the goal and an unambiguous identification of engineered most cancers cells. Right here, we current a platform consisting of: (i) t(8;21) and t(15;17) pushed acute myeloid leukemia (AML) transgenic mice with constitutive expression of inexperienced fluorescent protein (GFP) and inducible expression of Cre recombinase, and (ii) REX, a modified pSico lentiviral vector for inducible shRNA expression and crimson fluorescent protein (RFP) as a range marker.
On this system, leukemic cells from transgenic mice are transduced with REX, circulate sorted, and transplanted into syngeneic hosts. Gene interference is induced in established tumors by tamoxifen remedy. Twin-color cell fluorescence guides the in vivo identification of shRNA interfered AML cells, monitoring engraftment and illness development. We examined the platform by inducing knockdown of Zeb2, a gene upregulated by AML1-ETO and PML-RARα oncogenes in pre-leukemic hematopoietic stem cell compartment, and noticed a big delay in leukemia onset. This proves the ability and utility of the platform and confirms Zeb2 contribution to the pathogenesis of AML. Transgenes are susceptible to progressive silencing as a result of their construction, copy quantity, and genomic location. In C. elegans, repressive mechanisms are significantly robust within the germline with nearly totally penetrant transgene silencing in easy extrachromosomal arrays and frequent silencing of single-copy transgene insertions.
A category of non-coding DNA, Periodic An/Tn Clusters (PATCs) can forestall transgene-silencing in repressive chromatin or from small interfering RNAs (piRNAs). Right here, we describe design guidelines (codon-optimization, intron and PATC inclusion, elevated temperature (25 °C), and vector spine elimination) for environment friendly germline expression from arrays in wildtype animals.
BDNF Overexpression Will increase Striatal D3 Receptor Degree at Striatal Neurons and Exacerbates D1-Receptor Agonist-Induced Dyskinesia
Background: We just lately confirmed that striatal overexpression of mind derived neurotrophic issue (BDNF) by adeno-associated viral (AAV) vector exacerbated L-DOPA-induced dyskinesia (LID) in 6-OHDA-lesioned rats. An intensive sprouting of striatal serotonergic terminals accompanied this impact, accounting for the elevated susceptibility to LID.
Goal: We set to research whether or not the BDNF impact was restricted to LID, or prolonged to dyskinesia induced by direct D1 receptor agonists.
Strategies: Unilaterally 6-OHDA-lesioned rats obtained a striatal injection of an AAV vector to induce BDNF overexpression. Eight weeks later, animals obtained every day remedies with a low dose of SKF82958 (0.02 mg/kg s.c.) and growth of dyskinesia was evaluated. On the finish of the experiment, D1 and D3 receptors expression ranges and D1 receptor-dependent signaling pathways have been measured within the striatum.
Outcomes: BDNF overexpression induced important worsening of dyskinesia induced by SKF82958 in comparison with the GFP group and elevated the expression of D3 receptor at striatal degree, even in absence of pharmacological remedy; in contrast, D1 receptor ranges weren’t affected. In BDNF-overexpressing striata, SKF82958 administration resulted in elevated ranges of D1-D3 receptors co-immunoprecipitation and elevated phosphorylation ranges of Thr34 DARPP-32 and ERK1/2.
Conclusion: Right here we offer proof for a purposeful hyperlink between BDNF, D3 receptors and D1-D3 receptor shut interplay within the augmented susceptibility to dyskinesia in 6-OHDA-lesioned rats. We propose that D1/D3 receptors interplay could also be instrumental in driving the molecular alterations underlying the looks of dyskinesia; its disruption could also be a therapeutic technique for treating dyskinesia in PD sufferers.
A Novel Platform to Test In Vivo Single Gene Dependencies in t(8,21) and t(15,17) AML Confirms Zeb2 as Leukemia Target
Agrobacterium-Mediated Genetic Transformation of Wild Oryza Species Utilizing Immature Embryos
Genetic transformation is likely one of the most essential applied sciences for revealing or modulating gene perform. It’s used extensively in each purposeful genomics and molecular breeding of rice. Calls for on its use in wild Oryza species is rising due to their excessive genetic range. Given the difficulties in genetic crosses between distantly associated species, genetic transformation provides a option to alter or switch genetic traits in wild rice accessions. Nevertheless, transformation of untamed Oryza accessions by typical strategies utilizing calli induced from scutellum tissue of embryos in mature seeds usually fails.
Right here, we report strategies utilizing immature embryos for the genetic transformation of a broad vary of Oryza species. First, we investigated the power of callus induction and regeneration from immature embryos of 192 accessions in 20 species below a number of tradition situations. We regenerated vegetation from immature embryos of 90 accessions in 16 species. Subsequent, we optimized the situations of Agrobacterium an infection utilizing a vector carrying the GFP gene pushed by the maize ubiquitin promoter. GFP alerts have been noticed in 51 accessions in 11 species. We analyzed the expansion and seed set of transgenic vegetation of O. barthii, O. glumaepatula, O. rufipogon, and O. brachyantha. The vegetation grew to maturity and set seeds usually. Southern blot analyses utilizing DNA from T0 vegetation confirmed that every one GFP vegetation have been derived from impartial transformation occasions.
Express Cloning Checker Kit I: Yellow Solution |
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| K5011200-2 | Biochain | 1 ml | EUR 68 |
Checker Plus pH Tester |
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| HI98100 | Scientific Laboratory Supplies | EACH | EUR 72 |
SLS Lab Basics pH Checker |
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| PHM9500 | Scientific Laboratory Supplies | EACH | EUR 66.59 |
Mut Express II Fast Mutagenesis Kit V2 |
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| C214-01 | Vazyme | 10 rxn | EUR 198 |
Mut Express II Fast Mutagenesis Kit V2 |
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| C214-02 | Vazyme | 25 rxn | EUR 283.2 |
pCDF1-MCS1 cDNA Cloning and Expression Vector |
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| CD100A-1 | SBI | 10 ug | EUR 483 |
pCDH-CMV-MCS cDNA Cloning and Expression Vector |
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| CD500B-1 | SBI | 10 ug | EUR 483 |
pCDH-EF1-MCS cDNA Cloning and Expression Vector |
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| CD502A-1 | SBI | 10 ug | EUR 483 |
pCDH-CMV-MCS2 cDNA Cloning and Expression Vector |
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| CD501A-1 | SBI | 10 ug | EUR 483 |
pSIH1-H1-Puro shRNA Cloning and Expression Vector |
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| SI500A-1 | SBI | 10 ug | EUR 507 |
pSIH1-H1-H2Kk shRNA Cloning and Expression Vector |
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| SI502A-1 | SBI | 10 ug | EUR 507 |
pCDH-EF1-MCS-(PGK-GFP) Cloning and Expression Vector |
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| CD811A-1 | SBI | 10 ug | EUR 579 |
pAAVK-EF1α -MCS AAVanced Cloning and Expression Vector |
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| AAV502A-1 | SBI | 10 ug | EUR 614 |
pSIH1-H1-copGFP shRNA Cloning and Expression Vector |
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| SI501A-1 | SBI | 10 ug | EUR 507 |
pCDH-EF1-MCS-(PGK-Puro) Cloning and Expression Vector |
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| CD810A-1 | SBI | 10 ug | EUR 579 |
Hanna Checker pH Tester HI 98103 with 0.1 pH Resolution |
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| PHM4000 | Scientific Laboratory Supplies | EACH | EUR 78 |
Cumate-Inducible (CuO) EEV cloning and expression vector |
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| EEV610A-1 | SBI | 10 ug | EUR 614 |
PB-CMV-MCS-EF1-GFP cDNA cloning and expression vector |
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| PB511B-1 | SBI | 10 ug | EUR 638 |
PB-CMV-MCS-EF1-RFP cDNA cloning and expression vector |
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| PB512B-1 | SBI | 10 ug | EUR 638 |
PB-CMV-MCS-EF1-Puro cDNA cloning and expression vector |
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| PB510B-1 | SBI | 10 ug | EUR 638 |
PB-EF1-MCS-IRES-GFP cDNA cloning and expression vector |
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| PB530A-2 | SBI | 10 ug | EUR 638 |
PB-EF1-MCS-IRES-RFP cDNA Cloning and Expression Vector |
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| PB531A-2 | SBI | 10 ug | EUR 638 |
PB-EF1-MCS-IRES-Neo cDNA cloning and expression vector |
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| PB533A-2 | SBI | 10 ug | EUR 638 |
Constitutive (CAGs promoter) EEV cloning and expression vector |
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| EEV600A-1 | SBI | 10 ug | EUR 614 |
pCDH-CMV-MCS-EF1-RFP cDNA Cloning and Expression Vector |
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| CD512B-1 | SBI | 10 ug | EUR 572 |
pCDH-CMV-MCS-EF1-Neo cDNA Cloning and Expression Vector |
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| CD514B-1 | SBI | 10 ug | EUR 572 |
pCDH-MCS-T2A-copGFP-MSCV Cloning and Expression Vector |
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| CD523A-1 | SBI | 10 ug | EUR 590 |
pCDF1-MCS2-EF1-copGFP cDNA Cloning and Expression Vector |
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| CD111B-1 | SBI | 10 ug | EUR 572 |
pCDH-CMV-MCS-EF1-Puro cDNA Cloning and Expression Vector |
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| CD510B-1 | SBI | 10 ug | EUR 572 |
pCDH-EF1-MCS-IRES-GFP cDNA Cloning and Expression Vector |
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| CD530A-2 | SBI | 10 ug | EUR 590 |
pCDH-EF1-MCS-IRES-RFP cDNA Cloning and Expression Vector |
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| CD531A-2 | SBI | 10 ug | EUR 590 |
pCDH-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector |
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| CD533A-2 | SBI | 10 ug | EUR 590 |
pCDH-UbC-MCS-IRES-GFP cDNA Cloning and Expression Vector |
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| CD630A-1 | SBI | 10 ug | EUR 590 |
pCDH-MSCV-MCS-EF1-GFP cDNA Cloning and Expression Vector |
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| CD711B-1 | SBI | 10 ug | EUR 572 |
pCDH-CMV-MCS-EF1-Hygro cDNA Cloning and Expression Vector |
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| CD515B-1 | SBI | 10 ug | EUR 572 |
pCDH-MCS-T2A-Puro-MSCV cDNA Cloning and Expression Vector |
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| CD522A-1 | SBI | 10 ug | EUR 590 |
pCDH-UbC-MCS-EF1-Hygro cDNA Cloning and Expression Vector |
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| CD615B-1 | SBI | 10 ug | EUR 572 |
pCDH-EF1-MCS-IRES-Puro cDNA Cloning and Expression Vector |
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| CD532A-2 | SBI | 10 ug | EUR 590 |
pCDH-MSCV-MCS-EF1-Puro cDNA Cloning and Expression Vector |
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| CD710B-1 | SBI | 10 ug | EUR 572 |
pCDH-MSCV-MCS-IRES-GFP cDNA Cloning and Expression Vector |
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| CD731B-1 | SBI | 10 ug | EUR 590 |
PB-CMV-MCS-EF1-RedPuro cDNA Cloning and Expression Vector |
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| PB514B-2 | SBI | 10 ug | EUR 638 |
pCDH-CMV-MCS-EF1-copGFP cDNA Cloning and Expression Vector |
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| CD511B-1 | SBI | 10 ug | EUR 572 |
pCDH-CMV-MCS-t2A-copGFP cDNA Cloning and Expression Vector |
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| CD524A-1 | SBI | 10 ug | EUR 590 |
pCDF1-MCS2-EF1-Puro cDNA Cloning and Expression Lentivector |
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| CD110B-1 | SBI | 10 ug | EUR 572 |
pCDH-CMV-MCS-EF1-RFP+Puro cDNA Cloning and Expression Vector |
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| CD516B-2 | SBI | 10 ug | EUR 572 |
pCDH-EF1-MCS-T2A-Puro cDNA Cloning and Expression Vector 10 µg |
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| CD527A-1 | SBI | 10 ug | EUR 590 |
PB-CMV-GreenPuro-H1-MCS shRNA cloning and expression vector |
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| PBSI505A-1 | SBI | 10 ug | EUR 638 |
PB-EF1-GreenPuro-H1-MCS shRNA cloning and expression vector |
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| PBSI506A-1 | SBI | 10 ug | EUR 638 |
PB-CMV-MCS-EF1-GreenPuro cDNA cloning and expression vector |
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| PB513B-1 | SBI | 10 ug | EUR 638 |
pCDH-EF1-MCS-T2A-RFP (PGK-Puro) Cloning and Expression Vector |
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| CD822A-1 | SBI | 10 ug | EUR 579 |
pCDH-EF1-MCS-T2A-GFP (PGK-Puro) Cloning and Expression Vector |
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| CD823A-1 | SBI | 10 ug | EUR 579 |
pCDH-MSCV-MCS-EF1-GFP+Puro cDNA Cloning and Expression Vector |
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| CD713B-1 | SBI | 10 ug | EUR 572 |
pCDH-EF1-MCS-(PGK-GFP-T2A-Puro) Cloning and Expression Vector |
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| CD813A-1 | SBI | 10 ug | EUR 579 |
pAAVK-EF1α -MCS-T2A-EGFP AAVanced Cloning and Expression Vector |
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| AAV526A-1 | SBI | 10 ug | EUR 614 |
pAAVK-EF1α -MCS-T2A-Puro AAVanced Cloning and Expression Vector |
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| AAV527A-1 | SBI | 10 ug | EUR 614 |
pAAVK-EF1α –MCS-T2A-mRFP AAVanced Cloning and Expression Vector |
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| AAV528A-1 | SBI | 10 ug | EUR 614 |
PB-MSCV-MCS-EF1-GreenPuro cDNA Cloning and Expression Vector |
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| PB713B-1 | SBI | 10 ug | EUR 638 |
pCDH-CMV-MCS-EF1-GreenPuro cDNA Cloning and Expression Vector |
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| CD513B-1 | SBI | 10 ug | EUR 572 |
pAAVK-EF1α-MCS1-CMV-MCS2 AAVanced Cloning and Expression Vector |
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| AAV503A-1 | SBI | 10 ug | EUR 614 |
pAAVK-EF1α-MCS1-CMV-EGFP AAVanced Cloning and Expression Vector |
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| AAV536A-1 | SBI | 10 ug | EUR 614 |
pAAVK-EF1α-MCS1-CMV-Puro AAVanced Cloning and Expression Vector |
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| AAV537A-1 | SBI | 10 ug | EUR 614 |
pAAVK-EF1α-MCS1-CMV-mRFP AAVanced Cloning and Expression Vector |
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| AAV538A-1 | SBI | 10 ug | EUR 614 |
Jade Express? Mix |
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| M1109-500 | Biovision | each | EUR 529.2 |
Staph Express Disk |
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| FSA1044 | Scientific Laboratory Supplies | PK100 | EUR 134.48 |
Staph Express Disk |
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| FSA1046 | Scientific Laboratory Supplies | PK20 | EUR 50.4 |
pCDH-EF1-MCS-BGH-PGK-GFP-T2A-Puro cDNA Cloning and Expression Vector |
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| CD550A-1 | SBI | 10 ug | EUR 563 |
Express CRISPR sgRNA Synthesis Kit |
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| K1253-25 | Biovision | each | EUR 639.6 |
XStamp cloning and expression lentivector MSCV-Leader-MCS-C1C2-EF1-Puro |
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| XSTP710PA-1 | SBI | 10 ug | EUR 730 |
PB-CuO-CMV-MCS-EF1α-CymR-Puro Inducible cDNA Cloning and Expression Vector |
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| PBQM800A-1 | SBI | 10 µg | EUR 921 |
Express Plasmid Midiprep Kit (25 min) |
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| K1323-10 | Biovision | each | EUR 411.6 |
Express Plasmid Midiprep Kit (25 min) |
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| K1323-2 | Biovision | each | EUR 314.4 |
Express Plasmid Midiprep Kit (25 min) |
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| K1323-25 | Biovision | each | EUR 561.6 |
Express Plasmid Maxiprep Kit (25 min) |
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| K1324-10 | Biovision | each | EUR 510 |
Express Plasmid Maxiprep Kit (25 min) |
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| K1324-2 | Biovision | each | EUR 339.6 |
Express Plasmid Maxiprep Kit (25 min) |
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| K1324-25 | Biovision | each | EUR 732 |
Jade Express? Mix-ROX |
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| M1112-500 | Biovision | each | EUR 529.2 |
PB-Cuo-MCS-IRES-GFP-EF1-CymR-Puro Inducible cDNA Cloning and Expression Vector |
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| PBQM812A-1 | SBI | 10 ug | EUR 1023 |
Staph Express Count Plate |
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| FSA1040 | Scientific Laboratory Supplies | PK500 | EUR 828 |
Staph Express Count Plate |
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| FSA1042 | Scientific Laboratory Supplies | PK50 | EUR 117.6 |
pDsRed- Express- C1 Plasmid |
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| PVT1236 | Lifescience Market | 2 ug | EUR 319.2 |
MILLIPAK EXPRESS 40 FILTER |
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| MPGP04001SIG | Scientific Laboratory Supplies | EACH | EUR 261.12 |
Jade Express? Mix-iCycler |
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| M1110-500 | Biovision | each | EUR 529.2 |
ToxOut? Express Endofree Plasmid Midi Kit |
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| K1335-10 | Biovision | each | EUR 444 |
ToxOut? Express Endofree Plasmid Midi Kit |
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| K1335-2 | Biovision | each | EUR 280.8 |
ToxOut? Express Endofree Plasmid Midi Kit |
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| K1335-25 | Biovision | each | EUR 836.4 |
ToxOut? Express Endofree Plasmid Maxi Kit |
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| K1336-10 | Biovision | each | EUR 574.8 |
ToxOut? Express Endofree Plasmid Maxi Kit |
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| K1336-2 | Biovision | each | EUR 314.4 |
ToxOut? Express Endofree Plasmid Maxi Kit |
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| K1336-25 | Biovision | each | EUR 1032 |
PB-Cuo-shMCS-IRES-GFP-EF1-CymR-Puro Inducible shRNA Cloning and Expression Vector |
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| PBQMSH812A-1 | SBI | 10 ug | EUR 1023 |
pIRES2- DsRed- Express Plasmid |
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| PVT1229 | Lifescience Market | 2 ug | EUR 289.2 |
Jade Express? Mix-Low ROX |
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| M1111-500 | Biovision | each | EUR 529.2 |
Star 1kb DNA Ladder Express |
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| BT10701 | Bioatlas | 500µl | EUR 96 |
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Description: High purity buffer for various PCR applications. |
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Mut Express MultiS Fast Mutagenesis Kit V2 |
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| C215-01 | Vazyme | 10 rxn | EUR 297.6 |
Mut Express MultiS Fast Mutagenesis Kit V2 |
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| C215-02 | Vazyme | 25 rxn | EUR 490.8 |
Cryo Express Dry Shipper (CX100) |
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| TW-CX100 | MiTeGen | 1 SHIPPER | EUR 1265 |
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Description: Cryo Express Dry Shipper (CX100) |
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Star 100 bp DNA Ladder Express |
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| BA01302 | Bioatlas | 500µl | EUR 96 |
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Description: High purity buffer for various PCR applications. |
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Millipak Express 20 Filter (1/box) |
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| MPGP02001 | Scientific Laboratory Supplies | EACH | EUR 279.4 |
BrightGreen Express 2X qPCR MasterMix-ROX |
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| MasterMix-ER | ABM | 4 x 1.25 ml for 500 reactions (20 ul) | EUR 168 |
BrightGreen Express 2X qPCR MasterMix-iCycler |
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| MasterMix-EC | ABM | 4 x 1.25 ml for 500 reactions (20 ul) | EUR 168 |
BrightGreen Express 2X qPCR MasterMix-No Dye |
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| MasterMix-ES | ABM | 4 x 1.25 ml for 500 reactions (20 ul) | EUR 168 |
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 168 |
ClonExpress II One Step Cloning Kit |
|||
| C112-01 | Vazyme | 25 rxn | EUR 202.8 |
ClonExpress II One Step Cloning Kit |
|||
| C112-02 | Vazyme | 50 rxn | EUR 259.2 |
Cryo Express Dry Shipper with replaceable absorbent Material (CXR100) |
|||
| TW-CXR100 | MiTeGen | 1 SHIPPER | EUR 1365 |
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Description: Cryo Express Dry Shipper with replaceable absorbent Material (CXR100) |
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Cryo Express Dry Shipper (CX100) with NEW Upright Shipping Case (4LSC) |
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| TW-CXS-2 | MiTeGen | 1 SHIPPER AND CASE | EUR 1703 |
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Description: Cryo Express Dry Shipper (CX100) with NEW Upright Shipping Case (4LSC) |
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T1 Cloning Kit |
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| 20-abx098055 | Abbexa |
|
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T3 Cloning Kit |
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| 20-abx098057 | Abbexa |
|
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EXPH5 cloning plasmid |
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| CSB-CL843342HU-10ug | Cusabio | 10ug | EUR 3324 |
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Description: A cloning plasmid for the EXPH5 gene. |
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Blunt Cloning Kit |
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| 20-abx098059 | Abbexa |
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Blunt3 Cloning Kit |
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| 20-abx098061 | Abbexa |
|
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Gryphon Retroviral Expression System II |
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| ABP-RVK-10002 | Allele Biotech | 1 set | Ask for price |
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Description: PRODUCT LINE DISCONTINUED. Please contact us. |
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CHEK2 cloning plasmid |
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| CSB-CL005339HU-10ug | Cusabio | 10ug | EUR 451.2 |
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Description: A cloning plasmid for the CHEK2 gene. |
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CHERP cloning plasmid |
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| CSB-CL005340HU-10ug | Cusabio | 10ug | EUR 1023.6 |
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Description: A cloning plasmid for the CHERP gene. |
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T5 Zero Cloning Kit |
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| 20-abx098058 | Abbexa |
|
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T1 Simple Cloning Kit |
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| 20-abx098056 | Abbexa |
|
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Blunt Zero Cloning Kit |
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| 20-abx098062 | Abbexa |
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Blunt Simple Cloning Kit |
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| 20-abx098060 | Abbexa |
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Dog Thymus Expressed Chemokine ELISA kit |
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| E08T0114-192T | BlueGene | 192 tests | EUR 1524 |
|
Description: A competitive ELISA for quantitative measurement of Canine Thymus Expressed Chemokine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Dog Thymus Expressed Chemokine ELISA kit |
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| E08T0114-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
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Description: A competitive ELISA for quantitative measurement of Canine Thymus Expressed Chemokine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Dog Thymus Expressed Chemokine ELISA kit |
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| E08T0114-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
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Description: A competitive ELISA for quantitative measurement of Canine Thymus Expressed Chemokine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Rat Thymus Expressed Chemokine ELISA kit |
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| E02T0114-192T | BlueGene | 192 tests | EUR 1524 |
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Description: A competitive ELISA for quantitative measurement of Rat Thymus Expressed Chemokine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Rat Thymus Expressed Chemokine ELISA kit |
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| E02T0114-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
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Description: A competitive ELISA for quantitative measurement of Rat Thymus Expressed Chemokine in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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We confirmed that the T-DNAs have been transmitted to the following technology via the segregation of GFP alerts within the T1 technology. These outcomes present that many Oryza species could be remodeled by utilizing modified immature-embryo strategies. It will speed up the usage of wild Oryza accessions in molecular genetic analyses and molecular breeding.