[Construction of recombinant adenovirus vector pAdxsi-GFP-HIF containing hypoxia inducible factor gene and its expression in endothelial cells].
OBJECTIVE
To assemble a recombinant adenovirus (pAdxsi-GFP-HIF) encoding human hypoxia inducible issue 1 α gene (HIF-1 α) and to precise it in endothelial cells.
METHODS
HIF-1 α gene was obtained from human lung most cancers cell line A549, which was cultured in hypoxia situation, by RT-PCR. The HIF-1 α gene was subcloned into shuttle vector p Shuttle-CMV-EGFP at KpnI and BamHI websites. After recognized with restriction enzymes, plasmid p Shuttle-GFP-HIF was linearized by digestion with restriction endonuclease I-CeuI and I-SceI, and subsequently cotransformed into E.coli DH5a with adenoviral spine plasmid pAdxsi to make homologous recombination.
After linearized by PacI, the homologous recombinant adenovirus plasmid was transfected into 293 cells to bundle and amplify. The recombinant adenovirus was contaminated with human umbilical vein endothelial cells (ECV304), and the expression degree of HIF-1 α protein was evaluated by ELISA.
RESULTS
The recombinant adenovirus vector containing HIF-1 α gene (pAdxsi-GFP-HIF) was efficiently constructed and amplified with titer of three.38 X 10(10) pfu/mL. The inexperienced fluorescence protein was detected below fluorescent microscope in ECV304 at 24h after transfection and with a stronger diploma after 48h. The focus of HIF-1 protein was (48.93 ±3.86)ng/mL in supernatant at 48 h after transfection.
CONCLUSIONS
A recombinant adenovirus vector pAdxsi-GFP-HIF, encoding human hypoxia inducible issue 1 α gene, has been constructed in vitro and expressed efficiently in ECV304 cells.
gfpvector
ProtoGlow ECL 200ml Kit |
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| NAT1010 | National Diagnostics | EACH | EUR 354 |
ProtoGlow ECL 500ml Kit |
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| NAT1012 | National Diagnostics | EACH | EUR 627.6 |
100G Acrylamide |
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| NAT1014 | National Diagnostics | 100G | EUR 120 |
500G Acrylamide |
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| NAT1016 | National Diagnostics | 500G | EUR 261.6 |
1KG Acrylamide |
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| NAT1018 | National Diagnostics | 1KG | EUR 412.8 |
25G AquaPor LE |
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| NAT1020 | National Diagnostics | 25G | EUR 132 |
100G AquaPor LE GTAC |
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| NAT1022 | National Diagnostics | 100G | EUR 250.8 |
500G AquaPor LE GTAC |
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| NAT1024 | National Diagnostics | 500G | EUR 878.4 |
25G AquaPor ES |
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| NAT1026 | National Diagnostics | 25G | EUR 152.4 |
100G AquaPor ES |
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| NAT1028 | National Diagnostics | 100G | EUR 350.4 |
25G AquaPor LM |
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| NAT1030 | National Diagnostics | 25G | EUR 288 |
100G AquaPor LM |
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| NAT1032 | National Diagnostics | 100G | EUR 860.4 |
25G AquaPor HR |
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| NAT1034 | National Diagnostics | 25G | EUR 236.4 |
100G AquaPor HR |
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| NAT1036 | National Diagnostics | 100G | EUR 592.8 |
25G AquaPor 3:1 |
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| NAT1038 | National Diagnostics | 25G | EUR 250.8 |
100G AquaPor 3:1 |
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| NAT1040 | National Diagnostics | 100G | EUR 570 |
25G Bis (Methylene Bis-Acrylamide) |
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| NAT1042 | National Diagnostics | 25G | EUR 100.8 |
25G DATD Ultrapure |
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| NAT1044 | National Diagnostics | 25G | EUR 168 |
250G Glycine |
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| NAT1046 | National Diagnostics | 250G | EUR 94.8 |
1KG Glycine Ultrapure |
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| NAT1048 | National Diagnostics | 1KG | EUR 128.4 |
250G Tris |
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| NAT1050 | National Diagnostics | 250G | EUR 102 |
1KG Tris Ultrapure |
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| NAT1052 | National Diagnostics | 1KG | EUR 177.6 |
100G Tricine |
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| NAT1054 | National Diagnostics | 100G | EUR 120 |
100G Ion Exchange Resin (mixed bed) |
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| NAT1056 | National Diagnostics | 100G | EUR 96 |
25G Riboflavin |
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| NAT1058 | National Diagnostics | 25G | EUR 96 |
25ML TEMED |
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| NAT1060 | National Diagnostics | 25ML | EUR 99.6 |
25G Ammonium Persulfate |
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| NAT1062 | National Diagnostics | 25G | EUR 96 |
100G Ammonium Persulfate |
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| NAT1064 | National Diagnostics | 100G | EUR 118.8 |
1G Dithiothreitol (DTT) |
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| NAT1066 | National Diagnostics | 1G | EUR 103.2 |
5G Dithiothreitol (DTT) |
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| NAT1068 | National Diagnostics | 5G | EUR 183.6 |
50ML 2-Mercaptoethanol |
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| NAT1070 | National Diagnostics | 50ML | EUR 112.8 |
100G SDS Ultrapure |
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| NAT1072 | National Diagnostics | 100G | EUR 111.6 |
1KG SDS Ultrapure |
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| NAT1074 | National Diagnostics | 1KG | EUR 457.2 |
250G Urea |
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| NAT1076 | National Diagnostics | 250G | EUR 98.4 |
1KG Urea |
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| NAT1078 | National Diagnostics | 1KG | EUR 138 |
450ML Glycerol Ultrapure |
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| NAT1080 | National Diagnostics | 450ML | EUR 132 |
200ML Tween 20 Ultrapure |
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| NAT1082 | National Diagnostics | 200ML | EUR 132 |
1L Tween 20 |
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| NAT1084 | National Diagnostics | 1L | EUR 169.2 |
200ML Formamide |
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| NAT1086 | National Diagnostics | 200ML | EUR 111.6 |
500ML Boric Acid |
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| NAT1088 | National Diagnostics | 500ML | EUR 103.2 |
100G EDTA |
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| NAT1090 | National Diagnostics | 100G | EUR 94.8 |
500G EDTA |
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| NAT1092 | National Diagnostics | 500G | EUR 177.6 |
GelDry Film 11 X 12 cm |
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| NAT1094 | National Diagnostics | PK50 | EUR 100.8 |
25ML Formamide CE |
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| NAT1096 | National Diagnostics | 25ML | Ask for price |
25ML Glass Bond |
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| NAT1098 | National Diagnostics | 25ML | EUR 158.4 |
450ML Glass Free |
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| NAT1100 | National Diagnostics | 450ML | EUR 196.8 |
GelDry Film 22.5 X 22.5 cm |
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| NAT1102 | National Diagnostics | PK50 | EUR 130.8 |
1L DEPC Treated Water |
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| NAT1104 | National Diagnostics | 1L | EUR 120 |
Sterling Rapid Silver Stain Kit |
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| NAT1106 | National Diagnostics | 1KIT | EUR 265.2 |
1L ProtoBlue Safe |
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| NAT1108 | National Diagnostics | 1L | EUR 159.6 |
4L ProtoBlue Safe |
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| NAT1110 | National Diagnostics | 4L | EUR 282 |
1L ProtoStain Blue |
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| NAT1112 | National Diagnostics | 1L | EUR 164.4 |
25ML Nuclistain |
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| NAT1114 | National Diagnostics | 25ML | EUR 128.4 |
450ML AcrylaGel 30% |
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| NAT1116 | National Diagnostics | 450ML | EUR 128.4 |
1L AcrylaGel 30% |
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| NAT1118 | National Diagnostics | 1L | EUR 177.6 |
450ML Bis-AcrylaGel 2% |
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| NAT1120 | National Diagnostics | 450ML | EUR 98.4 |
1L Bis-AcrylaGel 2% |
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| NAT1122 | National Diagnostics | 1L | EUR 111.6 |
450ML UreaGel 29:1 Concentrate |
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| NAT1124 | National Diagnostics | 450ML | EUR 133.2 |
1L UreaGel 29:1 Concentrate |
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| NAT1126 | National Diagnostics | 1L | EUR 183.6 |
UreaGel 29:1 System 1L Kit |
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| NAT1128 | National Diagnostics | EACH | EUR 171.6 |
UreaGel 29:1 System 2.2L Kit |
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| NAT1130 | National Diagnostics | EACH | EUR 253.2 |
450ML SequaGel UreaGel Concentrate |
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| NAT1134 | National Diagnostics | 1L | EUR 160.8 |
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| NAT1136 | National Diagnostics | 1KIT | EUR 171.6 |
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100ML SequaGel Buffer |
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| NAT1140 | National Diagnostics | 100ML | EUR 80.4 |
200ML SequaGel UreaGel Buffer |
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450ML SequaGel UreaGel-6 |
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| NAT1144 | National Diagnostics | 450ML | EUR 115.2 |
1L SequaGel UreaGel-6 |
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| NAT1146 | National Diagnostics | 1L | EUR 151.2 |
450ML SequaGel UreaGel-8 |
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450ML SequaGel UreaGel Diluent |
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90ML SequaGel Complete Buffer |
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200ML SequaGel UreaGel Complete Buffer |
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450ML SequaGel XR |
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1L SequaGel XR |
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100ML SequaGel XR Concentrate |
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Development of a brand new GFPvector and its use for Fusaruim oxysporum transformation.
On this examine, the gfp fragment as a reporter gene had built-in into the shape plasmid vector pBC-hygro which accommodates an expressive promoter of the fungus to facilitate the transformation of Fusarium oxysporum. The resultant plasmid pBC-hygro-GFP was recognized by digestion with enzymes. Binary plasmids pBC-hygro-GFP had been remodeled into F. oxysporum through the use of the PEG-CaCl2 mediated transformation approach. Outcomes present that the recombinant plasmid pBC-hygro-GFP was constructed appropriately.
The gfp gene was stably maintained and didn’t convey any vital lack of phenotype which might have an effect on the survival and behavior of the tagged strains. Introduction of the gfp gene into F. oxysporum gives a easy, particular and cost-effective technique of pressure identification for ecological research. Transcriptional reporter vectors had been constructed for utilizing the inexperienced fluorescent protein (GFP) reporter.
GFP co-expression reduces the A33R gene expression pushed by a fowlpox vector in replication permissive and non-permissive cell strains.
The event of an efficient prophylactic vaccine continues to be mandatory to enhance the security of the standard although-discontinued smallpox vaccine, and to guard from the specter of deliberate launch of variola virus. This want additionally arises from the variety of new instances of animal orthopoxvirus infections every year, and to cut back the danger to animal handlers.
Fowlpox (FP) recombinants solely replicate in avian species and have been developed in opposition to human infectious ailments, as they will elicit an efficient immune response, should not cross-reactive immunologically with vaccinia, and characterize safer and extra promising immunogens for immunocompromised people.
The intention of this examine was the characterisation of two new fowlpox recombinants expressing the A33R vaccinia virus gene both alone (FP(A33R)) or with the inexperienced fluorescent protein (FP(A33R-GFP)) to confirm whether or not GFP can have an effect on the expression of the transgene.
The outcomes present that each FP(A33R) and FP(A33R-GFP) can categorical A33R appropriately, however A33R mRNA and protein synthesis are increased by FP(A33R) than by FP(A33R-GFP). Due to this fact, GFP co-expression doesn’t stop, however can cut back the extent of a vaccine protein, and will have an effect on the protecting efficacy of the immune response
100G Acrylamide |
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1KG Acrylamide |
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25G AquaPor LE |
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500G AquaPor LE GTAC |
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25G AquaPor ES |
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| NAT1026 | National Diagnostics | 25G | EUR 152.4 |
100G AquaPor ES |
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| NAT1028 | National Diagnostics | 100G | EUR 350.4 |
25G AquaPor LM |
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| NAT1030 | National Diagnostics | 25G | EUR 288 |
100G AquaPor LM |
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| NAT1032 | National Diagnostics | 100G | EUR 860.4 |
25G AquaPor HR |
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| NAT1034 | National Diagnostics | 25G | EUR 236.4 |
100G AquaPor HR |
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| NAT1036 | National Diagnostics | 100G | EUR 592.8 |
25G AquaPor 3:1 |
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| NAT1038 | National Diagnostics | 25G | EUR 250.8 |
100G AquaPor 3:1 |
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| NAT1040 | National Diagnostics | 100G | EUR 570 |
25G Bis (Methylene Bis-Acrylamide) |
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| NAT1042 | National Diagnostics | 25G | EUR 100.8 |
25G DATD Ultrapure |
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| NAT1044 | National Diagnostics | 25G | EUR 168 |
250G Glycine |
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| NAT1046 | National Diagnostics | 250G | EUR 94.8 |
1KG Glycine Ultrapure |
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| NAT1048 | National Diagnostics | 1KG | EUR 128.4 |
250G Tris |
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| NAT1050 | National Diagnostics | 250G | EUR 102 |
1KG Tris Ultrapure |
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| NAT1052 | National Diagnostics | 1KG | EUR 177.6 |
100G Tricine |
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| NAT1054 | National Diagnostics | 100G | EUR 120 |
100G Ion Exchange Resin (mixed bed) |
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| NAT1056 | National Diagnostics | 100G | EUR 96 |
25G Riboflavin |
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| NAT1058 | National Diagnostics | 25G | EUR 96 |
25ML TEMED |
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| NAT1060 | National Diagnostics | 25ML | EUR 99.6 |
25G Ammonium Persulfate |
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| NAT1062 | National Diagnostics | 25G | EUR 96 |
100G Ammonium Persulfate |
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| NAT1064 | National Diagnostics | 100G | EUR 118.8 |
1G Dithiothreitol (DTT) |
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| NAT1066 | National Diagnostics | 1G | EUR 103.2 |
5G Dithiothreitol (DTT) |
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| NAT1068 | National Diagnostics | 5G | EUR 183.6 |
50ML 2-Mercaptoethanol |
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| NAT1070 | National Diagnostics | 50ML | EUR 112.8 |
100G SDS Ultrapure |
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| NAT1072 | National Diagnostics | 100G | EUR 111.6 |
1KG SDS Ultrapure |
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| NAT1074 | National Diagnostics | 1KG | EUR 457.2 |
250G Urea |
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| NAT1076 | National Diagnostics | 250G | EUR 98.4 |
1KG Urea |
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| NAT1078 | National Diagnostics | 1KG | EUR 138 |
450ML Glycerol Ultrapure |
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| NAT1080 | National Diagnostics | 450ML | EUR 132 |
200ML Tween 20 Ultrapure |
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| NAT1082 | National Diagnostics | 200ML | EUR 132 |
1L Tween 20 |
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| NAT1084 | National Diagnostics | 1L | EUR 169.2 |
200ML Formamide |
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| NAT1086 | National Diagnostics | 200ML | EUR 111.6 |
500ML Boric Acid |
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| NAT1088 | National Diagnostics | 500ML | EUR 103.2 |
100G EDTA |
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| NAT1090 | National Diagnostics | 100G | EUR 94.8 |
500G EDTA |
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| NAT1092 | National Diagnostics | 500G | EUR 177.6 |
GelDry Film 11 X 12 cm |
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| NAT1094 | National Diagnostics | PK50 | EUR 100.8 |
25ML Formamide CE |
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| NAT1096 | National Diagnostics | 25ML | Ask for price |
25ML Glass Bond |
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| NAT1098 | National Diagnostics | 25ML | EUR 158.4 |
450ML Glass Free |
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| NAT1100 | National Diagnostics | 450ML | EUR 196.8 |
GelDry Film 22.5 X 22.5 cm |
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| NAT1102 | National Diagnostics | PK50 | EUR 130.8 |
1L DEPC Treated Water |
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| NAT1104 | National Diagnostics | 1L | EUR 120 |
Sterling Rapid Silver Stain Kit |
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| NAT1106 | National Diagnostics | 1KIT | EUR 265.2 |
1L ProtoBlue Safe |
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| NAT1108 | National Diagnostics | 1L | EUR 159.6 |
4L ProtoBlue Safe |
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| NAT1110 | National Diagnostics | 4L | EUR 282 |
1L ProtoStain Blue |
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| NAT1112 | National Diagnostics | 1L | EUR 164.4 |
25ML Nuclistain |
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| NAT1114 | National Diagnostics | 25ML | EUR 128.4 |
450ML AcrylaGel 30% |
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| NAT1116 | National Diagnostics | 450ML | EUR 128.4 |
1L AcrylaGel 30% |
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| NAT1118 | National Diagnostics | 1L | EUR 177.6 |
450ML Bis-AcrylaGel 2% |
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1L Bis-AcrylaGel 2% |
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450ML UreaGel 29:1 Concentrate |
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1L UreaGel 29:1 Concentrate |
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UreaGel 29:1 System 1L Kit |
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UreaGel 29:1 System 2.2L Kit |
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450ML SequaGel UreaGel Concentrate |
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1L SequaGel UreaGel Concentrate |
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SequaGel Sequencing System 1L Kit |
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SequaGel Sequencing System 2.2L Kit |
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| NAT1138 | National Diagnostics | EACH | EUR 253.2 |
100ML SequaGel Buffer |
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200ML SequaGel UreaGel Buffer |
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450ML SequaGel UreaGel-6 |
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1L SequaGel UreaGel-6 |
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450ML SequaGel UreaGel-8 |
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1L SequaGel UreaGel-8 |
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| NAT1150 | National Diagnostics | 1L | EUR 151.2 |
450ML SequaGel UreaGel Diluent |
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| NAT1152 | National Diagnostics | 450ML | EUR 90 |
1L SequaGel UreaGel Diluent |
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| NAT1154 | National Diagnostics | 1L | EUR 103.2 |
90ML SequaGel Complete Buffer |
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| NAT1156 | National Diagnostics | 90ML | EUR 79.2 |
200ML SequaGel UreaGel Complete Buffer |
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| NAT1158 | National Diagnostics | 200ML | EUR 85.2 |
450ML SequaGel XR |
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| NAT1160 | National Diagnostics | 450ML | EUR 126 |
1L SequaGel XR |
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| NAT1162 | National Diagnostics | 1L | EUR 165.6 |
100ML SequaGel XR Concentrate |
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| NAT1164 | National Diagnostics | 100ML | EUR 128.4 |
450ML SequaGel XR Concentrate |
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| NAT1166 | National Diagnostics | 450ML | EUR 361.2 |
200ML SequaGel MD Monomer Solution |
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| NAT1168 | National Diagnostics | 200ML | EUR 220.8 |
SequaGel MD SSCP Kit |
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| NAT1170 | National Diagnostics | 1KIT | EUR 253.2 |
SequaGel MD Heteroduplex Kit |
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| NAT1172 | National Diagnostics | EACH | EUR 265.2 |
1.2ML SequaGel MD SSCP Stop Solution |
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| NAT1174 | National Diagnostics | 1.2ML | EUR 85.2 |
450ML AccuGel 19:1 (30%) |
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| NAT1176 | National Diagnostics | 450ML | EUR 126 |
1L AccuGel 19:1 (30%) |
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| NAT1178 | National Diagnostics | 1L | EUR 165.6 |
450ML AccuGel 19:1 (40%) |
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1L AccuGel 19:1 (40%) |
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450ML AccuGel 29:1 (30%) |
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| NAT1184 | National Diagnostics | 450ML | EUR 126 |