TargetAmpTM
RNA Amplification for Microarray
TargetAmp™ 2-Round Aminoallyl-aRNA Amplification Kit 1.0
TargetAmp™: Selection Guide | Process | Performance | FAQ | Poster (3 MB PDF)
Applications
The TargetAmp™ 2-Round Aminoallyl-aRNA Amplification Kit 1.0* generates at least 5-million-fold amplification of the poly(A) RNA contained in a total cellular RNA sample. This amplification is sufficient to generate microgram amounts of aminoallyl-labeled antisense RNA (AA-aRNA; also called AA-cRNA) from the total RNA contained in a single cell. The kit uses an Eberwine-type linear RNA amplification method,1 but incorporates a number of improvements that significantly increase the yields of AA-aRNA obtained, while reducing the level of nonspecific amplification products. The kit is recommended for amplifying mRNA in total RNA from 1 to ~50 cells.
The kit provides all reagents (except reverse transcriptases and RNA purification columns) for generating AA-aRNA. The entire procedure takes only ~4 hours of hands-on time and can be completed in 2 days. Multiple samples can be amplified simultaneously. Once produced, the aminoallyl-aRNA can be readily labeled using biotin-NHS or fluorescent dye-NHS compounds. Biotin-NHS is available from Epicentre.
- Amplification and aminoallyl-labeling of mRNA in total RNA of 1-50 cells (10-500 pg RNA) for use in microarray analysis. Ideal for small samples such as laser-capture microdissected cells, biopsy samples, cultured cells, or other minute cell or tissue samples.
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Figure 1. The TargetAmp™ Kit 1.0 amplifies RNA from a single cell. A: Laser capture of a single cell from a section of rat dorsal root ganglion (DRG). B: Agarose gel of the ~1-3 µg AA-aRNA obtained using the TargetAmp Kit 1.0 to amplify mRNA in the ~10 pg total RNA isolated from two different laser-captured DRG cells. C: Scatter plot of microarray results obtained when Cy™ dye-labeled aRNA target prepared using the TargetAmp Kit 1.0 was hybridized to a glass slide spotted with rat brain cDNA. |
The kit provides all reagents (except reverse transcriptases and RNA purification columns) for generating AA-aRNA. The entire procedure takes only ~4 hours of hands-on time and can be completed in 2 days. Multiple samples can be amplified simultaneously. Once produced, the aminoallyl-aRNA can be readily labeled using biotin-NHS or fluorescent dye-NHS compounds. Biotin-NHS is available from Epicentre.
Benefits
*TargetAmp™ aRNA Amplification Products are covered by intellectual property licensed to Epicentre Technologies Corporation from Johnson & Johnson Pharmaceutical Research & Development, L.L.C., and by intellectual property sublicensed to Epicentre Technologies Corporation from Incyte Corporation.
TargetAmp™ 2-Round aRNA Amplification Kit 2.0
- The kit generates microgram amounts of AA-aRNA from as little as 10 pg of total cellular RNA (Table 1 and Fig. 1), representing greater than 5,000,000-fold amplification.
- The entire two-round RNA amplification process can be completed in 2 days and requires only about 4 hours of hands-on time.
- Multiple RNA samples can be easily amplified simultaneously.
- Highly reproducible amplification and microarray results.
- Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
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Table 1. The TargetAmp™ Kit 1.0 produces microgram amounts of AA-aRNA from as little as 10 pg (one cell) of total RNA.Results represent the average of multiple experiments. a. Approximately 5% of rat brain total RNA is poly(A) RNA. b. Approximately 2% of HeLa total RNA is poly(A) RNA. c. Approximately 1.5%-2% of rat kidney total RNA is poly(A) RNA. |
TargetAmp™ 2-Round aRNA Amplification Kit 2.0
Applications
- Production of microgram amounts of unlabeled aRNA from mRNA in total RNA of 1-50 cells (10-500 pg RNA) for microarray-based analysis. Ideal for small samples such as laser-capture microdissected cells, biopsy samples, cultured cells, or other minute cell or tissue samples.
- Generation of amplified aRNA from small samples for use in RT-PCR studies, or for archiving valuable samples.
The kit provides all reagents (except reverse transcriptases and RNA purification columns) for generating unlabeled aRNA. Since each NTP is provided in the kit as a separate solution, researchers can also use published methods or develop new methods to make labeled aRNA by direct incorporation of labeled nucleotides during the final in vitro transcription step.
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Table 1. The TargetAmp™ Kit 2.0 produces high yields of unlabeled aRNA from as little as 10 pg of input total RNA. Results represent the average of multiple experiments. a. Approximately 5% of rat brain total RNA is poly(A) RNA.
b. Approximately 2% of HeLa total RNA is poly(A) RNA. c. Approximately 1.5%-2% of rat kidney total RNA is poly(A) RNA. |
Benefits
TargetAmp™ 2-Round Biotin-aRNA Amplification Kit 3.0
- The kit generates microgram amounts of unlabeled aRNA from as little as 10 pg of total cellular RNA (Table 1), representing greater than 5,000,000-fold amplification.
- The entire two-round RNA amplification process can be completed in 2 days and requires only about 4 hours of hands-on time.
- Multiple RNA samples can be easily amplified simultaneously.
- Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
TargetAmp™ 2-Round Biotin-aRNA Amplification Kit 3.0
Applications
The TargetAmp™ 2-Round Biotin-aRNA Amplification Kit 3.0* is the only kit that can produce microgram amounts of biotin-antisense RNA (biotin-aRNA; also called cRNA) from as little as five cells (about 50 pg of total RNA) for microarray studies. The kit uses an Eberwine-type linear RNA amplification method,1 but incorporates a number of improvements that significantly increase the yields of biotin-aRNA obtained, while reducing the level of nonspecific RNA amplification products.
The TargetAmp™ 2-Round Biotin-aRNA Amplification Kit 3.0 provides all reagents (except reverse transcriptases and RNA purification columns) to amplify input RNA from 5 to ~50 cells. The entire procedure takes only ~4 hours of hands-on time and can be completed in 2 days. Multiple samples can be amplified simultaneously.
- Amplification and biotin-labeling of mRNA in total RNA of 5-50 cells (50-500 pg RNA) for use in microarray analysis.
The TargetAmp™ 2-Round Biotin-aRNA Amplification Kit 3.0* is the only kit that can produce microgram amounts of biotin-antisense RNA (biotin-aRNA; also called cRNA) from as little as five cells (about 50 pg of total RNA) for microarray studies. The kit uses an Eberwine-type linear RNA amplification method,1 but incorporates a number of improvements that significantly increase the yields of biotin-aRNA obtained, while reducing the level of nonspecific RNA amplification products.
The TargetAmp™ 2-Round Biotin-aRNA Amplification Kit 3.0 provides all reagents (except reverse transcriptases and RNA purification columns) to amplify input RNA from 5 to ~50 cells. The entire procedure takes only ~4 hours of hands-on time and can be completed in 2 days. Multiple samples can be amplified simultaneously.
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Table 1. The TargetAmp™ Kit 3.0 reaction produces high yields of biotin-aRNA from as little as 50 pg of total cellular RNA.Results represent the average of multiple experiments. a. Approximately 2% of HeLa total RNA is poly(A) RNA. |
Benefits
- The kit generates microgram amounts of biotin-aRNA from as little as five cells (about 50 pg of input total cellular RNA; Table 1).
- Multiple RNA samples can be easily amplified and labeled simultaneously.
- Linear RNA amplification process ensures preservation of the transcript profile of the sample.
- Highly reproducible amplification, labeling, and microarray results (Table 2).
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Table 2. Biotin-aRNA produced by the TargetAmp™ Kit 3.0 allows high-sensitivity transcript detection. Biotin-aRNA was prepared from the specified amount of mouse liver and skeletal muscle total RNAs using the TargetAmp 2-Round Biotin-aRNA Amplification Kit 3.0 and then hybridized to the Illumina®MouseRef-8 BeadChip array in duplicates. Data were imported into BeadStudio (Illumina) for analysis. Genes called "detected" are those with detection p-value <0.05. |
- Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
For research use only. Some applications in which this product may be used may be covered by patents or patent applications applicable in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a license, depending upon the particular application and country in which the product is used.
TargetAmp™ 1-Round Aminoallyl-aRNA Amplification Kit 101
Applications
- Amplification and aminoallyl-labeling of mRNA in total RNA of >2,500 cells (≥25 ng RNA) for use in microarray analysis.
The kit provides all reagents (except reverse transcriptase and RNA purification columns) for generation of AA-aRNA. The entire procedure takes about 2 hours of hands-on time and can be completed in 1 day, even with multiple samples.
Once produced, the aminoallyl-aRNA can be readily labeled using biotin-NHS or fluorescent dye-NHS compounds. Biotin-NHS is available from Epicentre.
Once produced, the aminoallyl-aRNA can be readily labeled using biotin-NHS or fluorescent dye-NHS compounds. Biotin-NHS is available from Epicentre.
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Table 1. The TargetAmp™ Kit 101 produces high yields of AA-aRNA from as little as 25 ng of input RNA. Results represent the average of multiple experiments. a. Approximately 2% of HeLa total RNA is poly(A) RNA. |
Benefits
- The kit generates microagram amounts of AA-aRNA from as little as 25 ng of total cellular RNA (Table 1), representing greater than 5,000-fold amplification.
- The simple, single-tube protocol can be completed in 1 day and requires only about 2 hours of hands-on time.
- Multiple RNA samples can be easily amplified simultaneously.
- Microarray results using AA-aRNA produced with the TargetAmp Kit 101 are highly reproducible.
- The TargetAmp Kit 101 has been validated by the microarray group of a leading pharmaceutical company for aRNA yield, amplification efficiency, AA-aRNA size and quality, and reproducibility of transcript detection.
- Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
TargetAmp™ 1-Round aRNA Amplification Kit 103
Applications
- Production of microgram amounts of unlabeled aRNA from mRNA in total RNA of >2,500 cells (≥25 ng RNA).
- Generation of unlabeled aRNA for use in RT-PCR, or for other applications.
Benefits
References
*TargetAmp™ aRNA Amplification Products are covered by intellectual property licensed to Epicentre Technologies Corporation from Johnson & Johnson Pharmaceutical Research & Development, L.L.C., and by intellectual property sublicensed to Epicentre Technologies Corporation from Incyte Corporation.
TargetAmp™ 1-Round Biotin-aRNA Amplification Kit 105
- The kit generates microgram amounts of aRNA from as little as 25 ng of input total RNA (Table 1), representing greater than 5,000-fold amplification.
- Simple, single-tube protocol can be completed in 1 day and requires only about 2 hours of hands-on time.
- Provides flexible alternatives to aminoallyl-UTP labeling of aRNA, including incorporation of biotin-labeled or dye-labeled nucleotides, synthesis of labeled cDNA using aRNA as template for reverse transcription, or direct labeling with biotin for use with microarrays or other applications.
- Multiple RNA samples can be easily amplified simultaneously.
- The TargetAmp Kit 103 has been optimized for aRNA yield, amplification efficiency, and aRNA size and quality.
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Table 1. The TargetAmp™ Kit 103 produces high yields of unlabeled aRNA from as little as 25 ng of total RNA.Results represent the average of multiple experiments. a. Approximately 2% of HeLa total RNA is poly(A) RNA. |
References
- Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
*TargetAmp™ aRNA Amplification Products are covered by intellectual property licensed to Epicentre Technologies Corporation from Johnson & Johnson Pharmaceutical Research & Development, L.L.C., and by intellectual property sublicensed to Epicentre Technologies Corporation from Incyte Corporation.
TargetAmp™ 1-Round Biotin-aRNA Amplification Kit 105
Applications
- Amplification and biotin-labeling of mRNA in total RNA of >2,500 cells (≥25 ng RNA) for use in microarray analysis.
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Table 1. A TargetAmp™ Kit 105 reaction produces high yields of biotin-aRNA from as little as 25 ng of input total RNA.Results represent the average of multiple experiments. a. Approximately 2% of HeLa total RNA is poly(A) RNA. |
Benefits
References
For research use only. Some applications in which this product may be used may be covered by patents or patent applications applicable in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a license, depending upon the particular application and country in which the product is used.
- Simple, single-tube, 6-hour procedure. Prepare biotin-aRNA and begin hybridization the same day.
- Biotin-aRNA produced by the kit detected more transcripts (genes) than competitive kits and showed high correlation with MAQC TaqMan® data (Fig. 1).
- Multiple RNA samples can be easily amplified and labeled simultaneously.
- The kit generates microgram amounts of biotin-aRNA from as little as 25 ng of input total cellular RNA (Table 1).
- Linear RNA amplification process preserves the transcript profile of the sample.
- Highly reproducible amplification, labeling, and microarray results.
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Figure 1. Correlation of expression ratios obtained from qPCR and from microarrays hybridized to aRNA prepared with the TargetAmp™ 105 kit. X axis, log2 expression ratios determined from Human Genome U133 Plus 2.0 Arrays (Affymetrix) hybridized to targets prepared by the TargetAmp 105 kit; Y axis, log2 expression ratios determined by the MAQC, using TaqMan® assays. The high correlation obtained (r = 0.936) indicates the excellent fidelity of the amplification process. |
References
- Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
For research use only. Some applications in which this product may be used may be covered by patents or patent applications applicable in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a license, depending upon the particular application and country in which the product is used.
TargetAmp™-Nano Labeling Kit for Illumina® Expression BeadChip®
Note: This kit was formerly called the TargetAmp™ Nano-g™ Biotin-aRNA Labeling Kit for the Illumina® System.
The TargetAmp™-Nano Labeling Kit for Illumina® Expression BeadChip® produces micrograms of biotin-aRNA (also called cRNA) from as little as 25 ng of total RNA. The easy 6-hour reaction enables researchers to prepare biotin-aRNA and begin BeadChip hybridizations in the same day. Biotin-aRNA produced by the TargetAmp-Nano Kit provides sensitive gene detection and yields consistent differential gene expression results.
The TargetAmp™-Nano Labeling Kit for Illumina® Expression BeadChip® produces micrograms of biotin-aRNA (also called cRNA) from as little as 25 ng of total RNA. The easy 6-hour reaction enables researchers to prepare biotin-aRNA and begin BeadChip hybridizations in the same day. Biotin-aRNA produced by the TargetAmp-Nano Kit provides sensitive gene detection and yields consistent differential gene expression results.
Applications
Benefits
TargetAmp™-Pico Labeling Kit for Illumina® Expression BeadChip®
- Generation of biotin-aRNA target for use with Illumina® Expression BeadChip® arrays from as little as 25 ng total RNA.
- Sensitive gene detection.
- Easy, 6-hour procedure yields biotin-aRNA target and enables BeadChip hybridization in the same day.
- Linear RNA amplification process preserves the relative abundance of the original sample.
- High yields of biotin-aRNA from as little as 25 ng of total RNA makes the kit ideal for preparing replicates and archiving.
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Figure 1. Schematic overview of the TargetAmp™-Nano Kit procedure.The easy 6-hour reaction enables researchers to prepare biotin-aRNA target and begin BeadChip® hybridization in the same day. |
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Table 1. The TargetAmp™-Nano Labeling Kit for Illumina® Expression BeadChip® arrays produces microgram amounts of biotin-aRNA from as little as 25 ng of input total RNA.Results summarize experiments performed using the HeLa Total RNA control provided in the kit. |
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Figure 2. Genes detected using biotin-aRNA produced by the TargetAmp™-Nano Labeling Kit for Illumina® Expression BeadChip®. Biotin-aRNA was produced from the indicated amount and source of total RNA and hybridized to either the Illumina® MouseRef-8 Expression BeadChip® (mouse liver and mouse muscle RNA) or the HumanHT-12 v4 Expression BeadChip® (human ovary and human testicle). |
TargetAmp™-Pico Labeling Kit for Illumina® Expression BeadChip®
Application
Prepare biotin-antisense RNA (aRNA) for use with Illumina® Expression BeadChip® arrays from minute amounts of RNA. Ideal for samples such laser-capture microdissected (LCM) tissues, flow-sorted cells, or biopsy tissue.
The TargetAmp™-Pico Labeling Kit for Illumina® Expression BeadChip®* is the only kit that can produce micrograms amounts of biotin-aRNA (also called cRNA) from as little as 50 pg of total RNA for use on Illumina Expression BeadChip arrays. The kit uses a significantly improved Eberwine1 linear RNA amplification method, but incorporates a number of improvements that significantly increase the yields of biotin-aRNA obtained, while reducing the level of nonspecific RNA amplification products.
The TargetAmp™-Pico Labeling Kit for Illumina® Expression BeadChip®* is the only kit that can produce micrograms amounts of biotin-aRNA (also called cRNA) from as little as 50 pg of total RNA for use on Illumina Expression BeadChip arrays. The kit uses a significantly improved Eberwine1 linear RNA amplification method, but incorporates a number of improvements that significantly increase the yields of biotin-aRNA obtained, while reducing the level of nonspecific RNA amplification products.
Benefits
References
- Linear RNA amplification process ensures preservation of the transcript profile of the sample.
- High correlation with results obtained using the TargetAmp™-Nano Labeling Kit for Illumina Expression BeadChip.
- High yields of biotin-aRNA from as little as 50 pg of total RNA (Table 1).
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Table 1. The TargetAmp™-Pico Labeling Kit for Illumina® Expression BeadChip® arrays produces microgram amounts of biotin-aRNA from as little as 50 pg of input total RNA. Results summarize experiments performed using the HeLa Total RNA control provided in the kit. |
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Figure 1. Genes detected using biotin-aRNA produced by the TargetAmp™-Pico Labeling Kit for Illumina® Expression BeadChip®. Biotin-aRNA was produced from the indicated amount and source of total RNA and hybridized to the Illumina MouseRef-8 Expression BeadChip. |
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Figure 2. Correlation of expression ratios (click to enlarge). Mouse liver/skeletal muscle expression ratios were obtained with biotin-aRNA produced from 100 ng and 100 pg of total RNA using the TargetAmp-Nano™ and TargetAmp-Pico™ kits, respectively. |
References
Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.
*TargetAmp™ aRNA Amplification Products are covered by intellectual property licensed to EPICENTRE Technologies Corporation from Johnson & Johnson Pharmaceutical Research & Development, L.L.C., and by intellectual property sublicensed to EPICENTRE Technologies Corporation from Incyte Corporation.
For research use only. Some applications in which this product may be used may be covered by patents or patent applications applicable in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a license, depending upon the particular application and country in which the product is used.
RiboMultiplier™ Sense-RNA Amplification Kit
dsDNA Conversion Kit for RiboMultiplier™ sRNA
dsDNA Conversion Kit for RiboMultiplier™ sRNA
Applications
Production of sense RNA (sRNA) from 10-500 ng of total RNA. Use the amplified sRNA for:
Production of sense RNA (sRNA) from 10-500 ng of total RNA. Use the amplified sRNA for:
- Archiving valuable RNA samples for future use.
- Microarray studies requiring either sRNA target or labeled antisense cDNA target for improved 5´/3´ coverage.
- Real-time PCR studies of gene expression.
- Target for Roche-NimbleGen expression arrays.
- Preparation of templates for high-throughput (next-gen) sequencing.
- cDNA library construction.
Benefits
- sRNA shows excellent correlation with unamplified samples, indicating a faithful representation of the original gene expression profile (Fig. 2).
- The unique terminal-tagging procedure yields sRNA with an excellent 3´/5´ ratio and less 3´ bias compared to other oligo(dT)-primed RNA amplification processes (Table 1).
- Microgram amounts of sRNA are generated from as little as 10 ng of input total RNA (Table 2), with essentially no background amplification.
- The entire RiboMultiplier sRNA kit procedure can be completed in 1 day.
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Figure 1. Schematic overview of the RiboMultiplier Sense-RNA Amplification procedure. |
A. | B. |
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Figure 2. Correlation of differential gene expression data from sRNA produced by the RiboMultiplier™ Sense-RNA Amplification Kit and unamplified total RNA with MAQC TaqMan® assays (~1,000 genes). Plots of log2 gene expression ratios for human brain and universal human reference total RNA obtained for ds cDNA synthesized from either RiboMultiplier sRNA (Panel A, X-axis) or unamplified total RNA (Panel B, X-axis) versus corresponding known MAQC TaqMan ratios (Y-axis) showed a similarly high correlation value (r=0.92), demonstrating excellent fidelity of the amplified sRNA compared to the unamplified total RNA.
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Table 1. 3´/5´ ratios for RiboMultiplier™ sRNA compared to aRNA produced by another RNA amplification procedure.Random-primed cDNA samples prepared from either RiboMultiplier™ sRNA or T7-dT aRNA were used in qPCR, and the 3´/5´ sequence ratios were determined for several genes. In general, 3´/5´ ratios around 1.0 were seen for the RiboMultiplier sRNA sample, demonstrating the enhanced 5´ sequence representation in sRNA as compared to T7-dT aRNA. |
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Table 2. The RiboMultiplier™ sRNA Kit provides high yields of sRNA from as little as 10 ng of total RNA.
a. Approximately 2% of HeLa total RNA is poly(A) RN |