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Maintains the value of legacy data by the continued use of Enzo kit, the gold standard for GeneChip® visualization.
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Decreases experimental variation and standardizes biological data derived from microarrays with the universally accepted system.
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Maximizes incorporation efficiency and produces strong, clear displays by utilizing two biotin-labeled nucleotides and a high quality T7 polymerase.
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Convenient workflow with a flexible 4-16 hour transcription time and reagents supplied in a ready-to-use format.
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Enables correlation of results from experiment-to-experiment, project-to-project and lab-to-lab.
The Single-Round RNA Amplification and Biotin Labeling System provides an optimized protocol and reagents for the production of biotin-labeled antisense RNA (aRNA) from total cellular RNA samples in less than 24 hours for array analysis. This kit has been improved to yield sufficient aRNA quantity for most microarray platforms (10 µg minimum) with total RNA input of as low as 250 ng and as high as 5000 ng. The higher yield results from an improved cDNA purification system, now provided with the kit.
The Single-Round RNA Amplification and Biotin Labeling System has been optimized for superior performance with reduced variability and improved reproducibility and data quality. The complete system is composed of reagents for cDNA synthesis and purification and in vitro transcription labeling.
Note: Control RNA (100µg/ml) (Prod. no. ENZ-42406-CR) is available separately.
aRNA was generated from the old and new version of the kit in one experiment. Equal quantities of aRNA were hybridized to Affymetrix HU133 plus 2.0 expression arrays. Signal intensity obtained from the two arrays show a correlation value of 0.995.
The old and new versions of the Single-round RNA Amplification and Biotin Labeling System were used to amplify 250 and 500ng of Universal Human Reference RNA (Stratagene #740000). Reactions were performed in triplicate. The average yields are plotted with standard deviation shown as error bars.
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Product Details
| Quality Control: | - aRNA yield is ≥23µg
- aRNA purity range is between an A260/A280 ratio of 1.9 and 2.3
- Median aRNA size is equal to or greater than 1200 nt
- Affymetrix GeneChip criteria:
- % Present calls ≥ than 40%
- Scale factors ≤ 3.0
- 5’/3’ ratios for actin and GAPDH ~ 1.0 ± 0.3
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| Use/Stability: | Store Box A at -20˚C in a non-frost free freezer. Store Box B at ambient condition. |
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| Handling: | Avoid freeze/thaw cycles. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -20°C |
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| Contents: | Box A:
40 µl, dNTP Mix (dN)
10 µl, Promoter Primer (P)
20 µl, First Strand Buffer (FB)
80 µl, DTT (D)
10 µl, Reverse Transcriptase (RT)
10 µl, RNase Inhibitor (I)
50 µl, DNA Polymerase (DP)
10 µl, RNase H (RH)
150 µl, Second Strand Buffer (SB)
60 µl, IVT Reaction Buffer (RB)
60 µl, 10x Biotin-Labeled Ribonucleotides (B)
60 µl, Enhancer Cocktail (EC)
30 µl, T7 RNA Polymerase (T7)
2.5 mL, Nuclease-free Water (W)
Box B:
4.5 ml, cDNA Binding Buffer (BB)
1.2 ml, 5X Wash Solution (WS)
10 cDNA Purification Columns
10 cDNA Elution Tubes
10 Collection Tubes |
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| Technical Info/Product Notes: | Technical Note: A Performance Comparison of the Enzo Single Round RNA Amplification and Biotin Labeling System.
Best-in-Class performance: "Enzo kit is the best choice for routine Affymetrix GeneChip experiments."
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| Regulatory Status: | RUO - Research Use Only |
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Product Literature References
Changes in the Expression Profile of Pyroptosis-Related Genes in Senescent Retinal Pigment Epithelial Cells after Lutein Treatment: B.S. Mrozik, et al.; Curr. Issues Mol. Biol.
45 , 1500 (2023),
Abstract;
HELLS Is Negatively Regulated by Wild-Type P53 in Liver Cancer by a Mechanism Involving P21 and FOXM1: S. Schuller, et al.; Cancers
14, 459 (2022),
Abstract;
Astrocytes are direct cellular targets of lithium treatment: novel roles for lysyl oxidase and peroxisome-proliferator activated receptor-γ as astroglial targets of lithium: A.D. Rivera & A.M. Butt; Transl. Psychiatry
9, 211 (2019),
Abstract;
Full Text
Profiling of embryonic nuclear vs. cellular RNA in Arabidopsis thaliana: D. Slane, et al.; Genomics Data
4, 96 (2015),
Abstract;
Full Text
Brassinosteroid enhances resistance to fusarium diseases of barley: S.S. Ali, et al.; Phytopathology
103, 1260 (2013),
Abstract;
Full Text
Gene transcripts associated with BMI in the motor cortex and caudate nucleus of calorie restricted rhesus monkeys: A.C. Mitchell, et al.; Genomics
99, 144 (2012),
Abstract;
Full Text
Physical activity-associated gene expression signature in nonhuman primate motor cortex: A.C. Mitchell, et al.; Obesity
20, 692 (2012),
Abstract;
Differential gene expression in oligodendrocyte progenitor cells, oligodendrocytes and type II astrocytes: J.G. Hu, et al.; Tohoku J. Exp. Med.
223, 161 (2011),
Abstract;
Transcriptional analysis of rat piriform cortex following exposure to the organophosphonate anticholinesterase sarin and induction of seizures: K.D. Spradling, et al.; J. Neuroinflamm.
8, 83 (2011),
Abstract;
Full Text
Transcriptional responses of the nerve agent-sensitive brain regions amygdala, hippocampus, piriform cortex, septum, and thalamus following exposure to the organophosphonate anticholinesterase sarin: K.D. Spradling, et al.; J. Neuroinflammation
8, 84 (2011),
Abstract;
Full Text
A novel statistical algorithm for gene expression analysis helps differentiate pregnane X receptor-dependent and independent mechanisms of toxicity: M.A. Mongan, et al.; PLoS One
5, e15595 (2010),
Abstract;
Full Text
Transcriptomic and innate immune responses to Yersinia pestis in the lymph node during bubonic plague: J.E. Cornet, et al.; Infect. Immun.
78, 5086 (2010),
Abstract;
Full Text
General Literature References
Amplified RNA synthesized from limited quantities of heterogeneous cDNA: R.N. Van Gelder, et al.; PNAS USA
87, 1663 (1990),
Abstract;
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