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Gold standard transcript labeling system maintains the value of legacy data
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Incorporation of two biotin nucleotides yields brighter signal, improving data from microarray experiments
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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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Decreases experimental variation and standardizes biological data derived from microarrays with the universally accepted system.
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Enables correlation of results from experiment-to-experiment, project -to-project and lab-to-lab.
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Production of large amounts of biotin-labeled RNA targets by in vitro transcription available separately with our BIOARRAY HIGHYIELD® RNA Labeling Kit (Prod. No. ENZ-42655)
The Enzo Life Sciences' 100-Reaction 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 (1µg-5µg) in less than 24 hours for array analysis. While cDNA quantities produced are sufficient for gene expression, manual sample preparation involving multiple enzymatic reactions and product purification steps are time consuming and error-prone. The 100-Reaction Single- Round RNA Amplification and Biotin Labeling System has been optimized for superior performance across several automated liquid handling platforms.
The result is a comprehensive system that reduces variability while subsequently improving reproducibility, data quality and throughput for automated genomic environments.
The complete system is composed of reagents for cDNA synthesis and in vitro transcription labeling and does not provide materials required for purification.
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.
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.
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Product Details
| Quality Control: | Performance Testing:
Biotin-labeled aRNA is generated from 500 ng of human reference total RNA following the procedures in the User Manual. Labeled aRNA is purified and yield determined by absorbance using a UV/Vis spectrophotometer. Determination of labeled aRNA transcript size is performed by capillary electrophoresis. Hybridized of aRNA to Affymetrix GeneChip HU133 2.0 Plus microarrays serves as a final functional test.
Minimum Specifications:
- aRNA yield must be > 23 µg.
- aRNA purify must range between an A260/A280 ratio of 1.9 and 2.3.
- aRNA size must be 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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| 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: | 400 µL, dNTP Mix (dN); 100 µL, Promoter Primer (P); 200 µL, First Strand Buffer (FB); 800 µL, DTT (D);100 µL, Reverse Transcriptase (RT); 100 µL, RNase Inhibitor (I); 500 µL, DNA Polymerase (DP); 100 µL, RNase H (RH); 1.5 mL, Second Strand Buffer (SB); 600 µL, IVT Reaction Buffer (RB); 600 µL, 10x Biotin-Labeled Ribonucleotides (B); 600 µL, Enhancer Cocktail (EC); 300 µL, T7 RNA Polymerase (T7); 13 mL, Nuclease-free Water (W) |
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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
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Profiling of embryonic nuclear vs. cellular RNA in Arabidopsis thaliana: D. Slane, et al.; Genomics Data
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Brassinosteroid enhances resistance to fusarium diseases of barley: S.S. Ali, et al.; Phytopathology
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Gene transcripts associated with BMI in the motor cortex and caudate nucleus of calorie restricted rhesus monkeys: A.C. Mitchell, et al.; Genomics
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Physical activity-associated gene expression signature in nonhuman primate motor cortex: A.C. Mitchell, et al.; Obesity
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Differential gene expression in oligodendrocyte progenitor cells, oligodendrocytes and type II astrocytes: J.G. Hu, et al.; Tohoku J. Exp. Med.
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Transcriptional analysis of rat piriform cortex following exposure to the organophosphonate anticholinesterase sarin and induction of seizures: K.D. Spradling, et al.; J. Neuroinflammation
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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),
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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
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Transcriptomic and innate immune responses to Yersinia pestis in the lymph node during bubonic plague: J.E. Corner, et al.; Infect. Immun.
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