Shop All RNA Amplification Kits

WT Expression Kit Invitrogen™

The WT Expression Kit is an RNA amplification kit that is designed to generate amplified sense-strand cDNA ready for fragmentation and labeling using the Affymetrix® GeneChip® WT Terminal Labeling Kit.

Features of the WT Expression Kit:

• Optimized for use with Affymetrix® GeneChip® Human, Mouse, and Rat 1.0 ST Arrays
• Allows RNA samples as small as 50 ng to be analyzed on Affymetrix® GeneChip® Arrays
• Supports a streamlined workflow that does not require a separate rRNA depletion step
• Is ideal for high-sensitivity expression profiling

Eliminates separate rRNA depletion step, preserves transcriptome coverage

The first-generation Affymetrix® amplification method requires depletion of ribosomal RNA (rRNA) from RNA samples for optimal exon-level analysis. Conversely, the Ambion® WT Expression Kit uses a novel reverse transcription (RT) priming method that eliminates the need for a separate rRNA depletion step. The kit includes RT primers designed using a proprietary oligonucleotide matching algorithm that eliminates primer sequences with homology to known ribosomal RNAs. The result is complete and unbiased coverage of the transcriptome, with rRNA amplification levels significantly lower than those of other methods.

Consistent results from less input RNA
With the Ambion® WT Expression Kit, samples as small as 50 ng of total RNA can be analyzed on Affymetrix® GeneChip® Human, Mouse, and Rat Exon and Gene 1.0 ST Arrays. Previous methods required 1 µg of total RNA—often difficult or impossible to obtain from limited sources such as stem cells or small tissue samples. The modest requirement for input RNA permits the analysis of rare samples and provides for more efficient and cost-effective experimentation for most sample types. To demonstrate the performance of the WT Expression Kit, RNA from three sample types (HeLa cells, and Microarray Quality Control (MAQC) A and B samples) was prepared in triplicate using either the Affymetrix® GeneChip® WT cDNA Synthesis and Amplification Kit or the WT Expression Kit, and analyzed on Human Exon 1.0 ST Arrays. Total RNA (1 µg) prepared by the Affymetrix® protocol underwent an rRNA-depletion step while just 50 ng total RNA (20-fold less) was prepared for microarray analysis using the WT Expression Kit. Both sets of samples were handled according to the manufacturers' recommendations. Direct correlation of log2 ratios for MAQC samples A and B was high, (r >0.94) at both the exon and transcript levels. Correlation of log2 ratios with gene expression data obtained using TaqMan® Gene Expression assays for real-time PCR was also high for both kits (see Product Bulletin for data).

Identify more differential expression with less RNA
In addition to its lower input RNA requirement, the WT Expression Kit provides a significant increase in sensitivity. A greater number of probe sets detected above background was obtained at the exon level with the WT Expression Kit as a result of an increased signal-to-noise ratio. A comparison of differential expression analysis results from the two methods indicated that, while differentially expressed genes and exons of MAQC A and MAQC B samples were similar between the two kits (78% and 89%, respectively), significantly more differential expression was observed on arrays hybridized with samples prepared using the WT Expression Kit. 499 more genes and 6,850 more exons were found to be differentially expressed in samples prepared using the Ambion® kit, compared to samples prepared with the Affymetrix® kit.

Note: For manual use only. Not for robotics.

Amino Allyl MessageAmp™ II-96 aRNA Amplification Kit Invitrogen™

The Ambion® Amino Allyl MessageAmp™ II-96 aRNA Amplification Kit includes reagents for high- throughput aRNA amplification with amimo allyl NTP incorporation. The kit includes sufficient reagents for 100 reactions; Cy™3 dye is not included.

• Consistently higher yields with magnetic bead- compared to filter-based purification
• Highly reproducible aRNA yields and size (CV less than 10%)
• Flexible format allowing RNA amplification with a partial or full 96-well plate
• Simple integration into robotic platforms

Improved First- and Second-strand cDNA Synthesis
Included in the kit is ArrayScript™ RT, a rationally engineered M-MLV reverse transcriptase, which produces equivalent or higher yields of full-length cDNA compared to other enzymes. The second-strand cDNA synthesis reaction has also been optimized to be compatible with the first-strand cDNA products generated with ArrayScript™ RT to allow maximal conversion of first-strand cDNA into full-length double-stranded cDNA templates.

Increased Labeling Efficiency of Your aRNA
The Amino Allyl MessageAmp™ aRNA Amplification Kit incorporates amino allyl NTP into the aRNA followed by the coupling of its reactive amino group to an NHS ester label (e.g., Cy™ or another dye). This strategy offers several advantages over the direct incorporation of labeled NTPs. Direct incorporation of labeled NTPs is inefficient and results in low yields and low specific activity aRNA and high costs. Unlike lableled NTPs, amino allyl–modified NTPs are incorporated almost as efficiently as unmodified NTPs and are much less expensive than the dye coupled NTPs.

Accessory Product
The 96-Well Magnetic-Ring Stand (SKU#AM10050) features 96 powerful ring-shaped magnets arranged to cradle each well of a 96-well plate to capture beads thoroughly and quickly. Captured magnetic beads will form a donut-shaped pellet with a large hole in the center. This capture pattern facilitates both supernatant removal and subsequent bead resuspension. The stand is suitable for high-throughput applications conducted manually with multichannel pipettors or with robotic liquid handlers.

Arcturus™ Paradise™ PLUS 1.5 Round Kit Applied Biosystems™

The ARCTURUS® Paradise® PLUS FFPE RNA Amplification Kit enables unprecedented gene expression analysis of millions of previously inaccessible formalin-fixed, paraffin-embedded (FFPE) tissue samples. Amplify RNA from as little as 5 ng of fixed RNA using reagents and methods that have been specially optimized to overcome the challenges of cross-linked FFPE templates (Figure 4).

Key product features:
• Integrate Seamlessly into Current Workflow
• Secure Downstream Success with the Paradise® QC Kit
Integrate Seamlessly into Current Workflow
The Paradise® PLUS Reagent System is compatible with all major microarray platforms. Robust and reproducible results are achieved using tools and techniques commonly employed in today’s research laboratories (Figures 1, 2 & 3).

Secure Downstream Success with the Paradise® QC Kit
Not all FFPE samples contain high quality RNA. Identify the best samples for your studies using a simple, real-time PCR research technique, and then proceed with confidence using these pre-qualified samples.

Integrated Systems for Microgenomics
The Paradise®PLUS Reagent System is validated as part of the ARCTURUS® Complete System for Microgenomics®, an integrated solution for utilizing small quantities of RNA for gene expression analysis, and features the ARCTURUSXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations. ARCTURUS®' Systems for Microgenomics enable accurate and sensitive microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

For Research Use Only. Not for use in diagnostics procedures.

Arcturus™ Paradise™ PLUS 2 Round Kit, amino allyl Applied Biosystems™

The ARCTURUS® Paradise® PLUS FFPE RNA Amplification Kit enables unprecedented gene expression analysis of millions of previously inaccessible formalin-fixed, paraffin-embedded (FFPE) tissue samples. Amplify RNA from as little as 5 ng of fixed RNA using reagents and methods that have been specially optimized to overcome the challenges of cross-linked FFPE templates (Figure 4). Incorporation of aminoallyl NTPs permit subsequent conjugation of activated dyes.

Key product features:
• Integrate Seamlessly into Current Workflow
• Secure Downstream Success with the Paradise® QC Kit

Integrate Seamlessly into Current Workflow
The Paradise® PLUS Reagent System is compatible with all major microarray platforms. Robust and reproducible results are achieved using tools and techniques commonly employed in today’s research laboratories (Figures 1, 2 & 3).

Secure Downstream Success with the Paradise® QC Kit
Not all FFPE samples contain high quality RNA. Identify the best samples for your studies using a simple, real-time PCR research technique, and then proceed with confidence using these pre-qualified samples.

Integrated Systems for Microgenomics
The Paradise® PLUS Reagent System is validated as part of the ARCTURUS® Complete System for Microgenomics®, an integrated solution for utilizing small quantities of RNA for gene expression analysis, and features the ARCTURUSXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations. ARCTURUS®' Systems for Microgenomics enable accurate and sensitive microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

For Research Use Only. Not for use in diagnostics procedures.

Arcturus™ RiboAmp™ HS PLUS 1.5 Round Kit Applied Biosystems™

The ARCTURUS® RiboAmp® HS PLUS RNA Amplification Kit enables the researcher to perform molecular analysis on RNA samples that, until now, have been too small for microarray analysis. The RiboAmp® HS PLUS Kit offers unique linear amplification from as little as 100 pg of total RNA. Using a proprietary, patent- pending process for high-efficiency, high-fidelity linear amplification, messenger RNA is amplified up to 1,000,000-fold in two rounds. The RiboAmp® HS PLUS Kit produces amplified antisense RNA (aRNA), ready for labeling and microarray hybridization or real-time PCR. The RiboAmp® HS PLUS Kit can be used to generate biologically relevant results with cDNA and oligonucleotide array platforms.

Key product features:
• Efficient application—amplify from as few as ten cells
• High fidelity—maintain original mRNA representation after two amplification rounds
• High reliability—obtain consistent results

Amplify from as Few as Ten Cells
The RiboAmp® HS PLUS RNA Amplification Kit allows researchers to perform microarray studies using total RNA from as few as ten microdissected cells. With the RiboAmp® HS PLUS Kit, 100-500 pg of total RNA or as few as 10-50 cells captured by Laser Capture Microdissection (LCM) generate enough amplified RNA for cDNA or oligonucleotide microarray studies. For best results, use the PicoPure® RNA Isolation Kit to optimize RNA recovery from small samples prior to amplification.

Ensure High Fidelity after Two Rounds
The RiboAmp® HS PLUS RNA amplification method maintains the original mRNA representation after two amplification rounds, enabling accurate gene expression profiles from ultra small samples. Comparison of microarray hybridization results between cDNA probes generated from total cellular RNA and from aRNA obtained after two rounds of amplification using the RiboAmp® HS PLUS Kit reveals very high concordance between differentially expressed genes. This demonstrates the amplification fidelity needed for relevant differential gene expression data.

Obtain Consistent Results
When working with ultra-small samples, amplification reproducibility between samples is particularly important. Data from multiple amplifications and hybridizations of the same starting material using the RiboAmp® HS PLUS RNA Kit correlate exceptionally well.

For Research Use Only. Not for use in diagnostics procedures.

MessageAmp™ Premier RNA Amplification Kit Invitrogen™

The MessageAmp™ Premier RNA Amplification Kit is the latest innovation for the preparation of RNA samples for microarray analysis. The kit builds upon the linear RNA amplification method and reagents developed for MessageAmp™ II Kits with enhancements that create a simplified workflow, improved performance, and more flexible RNA input requirements. All MessageAmp™ kits employ the proven methodology for RNA sample preparation and labeling based on the original T7 in vitro transcription (IVT) amplification technology, known as the Eberwine or reverse transcription-IVT (RT-IVT) method. The RT-IVT method is considered the gold standard for sample preparation in microarray-based expression profiling. It is well documented in the current scientific literature and was experimentally validated using TaqMan® RT-PCR. MessageAmp™ Premier Kit features:

• Lowest RNA input requirements for a single round of amplification: 20 ng total RNA yields enough biotin-modified aRNA for most microarray formats, with results equivalent to those from samples prepared from significantly more starting material using other RT-IVT kits
• Very flexible RNA input requirements: 20–500 ng total RNA, depending on the tissue or cell type
• Streamlined workflow: depending on the RNA input amounts, only a single day of processing time is required to complete a microarray experiment
• Simplified protocol and streamlined reagents
• Complete kit: everything needed to prepare biotin-modified aRNA is included
• Rigorous quality control that includes GeneChip® analysis
• T7 MEGAscript® IVT delivers up to 50,000-fold amplification
• Single-tube format
• Easy-to-use, high-recovery, magnetic-bead aRNA purification

Using the MessageAmp&trade Premier RNA Amplification Kit
Based on this proven enzymatic amplification and labeling strategy, the MessageAmp™ Premier procedure starts with a simple cDNA synthesis reaction using ArrayScript™ reverse transcriptase. No cleanup step is required after the RT; cDNA is used directly in a high yield IVT reaction using MEGAscript® technology. The IVT is configured to incorporate the modified nucleotide, biotin-UTP, into the aRNA synthesized. (In this protocol, the amplified RNA is referred to as aRNA; in the literature, it is also commonly called cRNA.) Once purified, the aRNA is suitable for use on microarray gene expression analysis platforms designed for biotin-modified antisense RNA samples.

MessageAmp™ II aRNA Amplification Kit Invitrogen™

The MessageAmp II aRNA Amplification Kit amplifies as little as 100 pg of input RNA for GeneChip analysis. Advantages of the MessageAmp II aRNA Amplification Kit include:

• Increased amplification—up to 5,000-fold
• Greater sensitivity due to increased yields of full-length cDNA
• GeneChip analysis with as little as 100 ng input RNA
• Efficient labeling produces high percent present calls and low 3'/5' ratios
• Includes ArrayScript RT, a specifically engineered M-MLV reverse transcriptase
• Optimized second strand cDNA synthesis reaction

System overview
Expression profiling using gene arrays has become the most widely used method for assessing gene expression on a genome-wide/scale. The primary limitation of this technology is the requirement for large amounts of RNA for array hybridization. This hurdle is usually overcome by using T7 RNA polymerase-driven transcription of cDNA (obtained from total RNA samples) to amplify sufficient amounts of cRNA. This amplification method is the basis of the MessageAmp II aRNA Amplification Kit.

An improved kit for cDNA amplification
Included in the kit is ArrayScript RT, a rationally engineered M-MLV reverse transcriptase, which produces equivalent or higher yields of full-length cDNA compared to other enzymes. The use of ArrayScript RT in conjunction with the optimization of the second-strand cDNA synthesis reaction results in increased conversion of mRNA into full-length, double-stranded cDNA templates. In addition, since the in vitro transcription (IVT) uses MEGAscript technology, the IVT incubation times are shortened. These improvements in the MessageAmp II kit also carry through to two-round reactions, facilitating robust amplification of total RNA from samples as low as 100 pg.

Accessory products
Bio-11-UTP (Cat. No. AM8450) and Bio-16-UTP (Cat. No. AM8451) can be used in MessageAmp aRNA amplification reactions for target labeling for GeneChip® analysis. The Bio-UTPs are confirmed free of nonspecific endonuclease activity, exonuclease activity, and RNase activity, and are functionally tested using the MessageAmp™ II aRNA Amplification Kit.

Arcturus™ RiboAmp™ PLUS 1.5 Round Kit Applied Biosystems™

The Arcturus® RiboAmp® PLUS RNA Amplification Kit permits the researcher to perform molecular analysis on RNA samples that were previously considered too small for microarray analysis. The RiboAmp® PLUS Kit offers unique linear amplification from as little as 1 ng of total RNA. Using a proprietary process for high-efficiency, high-fidelity linear amplification, messenger RNA is amplified up to 1000-fold in one synthesis round and up to 1,000,000-fold in two rounds. The RiboAmp® PLUS Kit produces amplified antisense RNA (aRNA), ready for labeling and microarray hybridization or real-time PCR. The RiboAmp Plus Kit can be used to generate biologically relevant results with cDNA and oligonucleotide array platforms. We also offer RiboAmp® HS PLUS Kits for smaller RNA quantities (Figure 1).

Key product features:
• High fidelity – maintain native expression levels
• Greater sensitivity – detect low-abundance mRNA
• High reliability – deliver reproducible amplified RNA for microarray experiments
• Speedy application – process samples faster
• Enhanced specifity – reveal differences in gene expression between cell types

Maintain Native Expression Levels
The RiboAmp® PLUS RNA Amplification Kit offers unprecedented high-fidelity RNA amplification even from microscopic samples. Comparison of microarray results from total cellular RNA and from aRNA amplified using the RiboAmp® PLUS Kit reveal very high concordance between differentially expressed genes. This demonstrates the RiboAmp® PLUS Kit's ability to deliver the amplification fidelity needed for reliable data analysis from small samples (Figure 2).

Detect Low-abundance mRNA
The RiboAmp® PLUS RNA Amplification Kit enables amplification of mRNA in all abundance classes. Using RT-PCR, low-, medium-, and high-abundance genes are readily detected within the amplified RNA population. Amplification of mRNA in all abundance classes ensures that differential gene expression patterns will be identified (Figure 3).

Deliver Reproducible Amplified RNA for Microarray Experiments
The RiboAmp® PLUS RNA Amplification Kit consistently delivers aRNA ready for labeling and hybridization to expression microarrays with minimal amplification-to-amplification variability. Microarray analysis of independent amplifications from the same source RNA shows excellent correlation as shown in Figures 4 and 5.

Process Samples Faster
The RiboAmp® PLUS Kit includes complete reagents and devices for template synthesis, transcription, and nucleic acid purification. Tedious vacuum concentrations are completely eliminated by incorporating optimized chemistries and by using Arcturus' novel, high-recovery MiraCol™ Purification Columns (Figure 6). There are significantly fewer pipetting steps because the sample is eluted directly into reaction tubes and reagents are conveniently pre-dispensed and pre-mixed. In one eight-hour workday, the RiboAmp® PLUS Kit enables amplification, labeling, and microarray hybridization set up. With a convenient overnight in vitro transcription, a second round of amplification can be completed by the following morning (Figure 7).

Reveal Differences in Gene Expression between Cell Types
Accurate gene expression analysis requires the analysis of specific cell types without interference from surrounding cells. Starting with these pure cell populations often means working with small samples. Special technologies are needed to overcome the challenges of handling these precious samples. The combination of LCM, RNA amplification, and microarray analysis reveals differential gene expression between cell types (Figures 8, 9 & 10).

Integrated Systems for Microgenomics
The Arcturus® Complete System for Microgenomics® offers an integrated solution for preparing small quantities of RNA for gene expression analysis. The System features the ArcturusXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations, the PicoPure® RNA Isolation Kit to extract and purify RNA, and the RiboAmp® PLUS Kit for linear amplification of RNA (Figure 11). Arcturus' Systems for Microgenomics enable accurate and sensitive microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

For Research Use Only. Not for use in diagnostics procedures.

MessageAmp™ II-Bacteria RNA Amplification Kit Invitrogen™

The MessageAmp™ II-Bacteria RNA Amplification Kit is for amplification of bacterial RNA for highly sensitive, whole-genome expression analysis. The kit uses an in vitro transcription (IVT)–mediated linear amplification method optimized for use with bacterial RNA. Advantages of the MessageAmp™ II-Bacteria RNA Amplification Kit:

Maximum sensitivity—amplify as little as 10 ng of mRNA or 100 ng total RNA; a 10- to 500-fold reduction in the input RNA needed to probe microarrays
Highest yield of full-length transcripts—includes ArrayScript™ RT which produces up to twice the amount of full-length cDNA as M-MLV, AMV, or SuperScript® II
Stringently reliable data—reactions are highly reproducible, with Pearson's Correlation Coefficients >0.995

Using the MessageAmp™ II-Bacteria RNA Amplification Kit
This kit is the only commercially available kit for amplification of prokaryotic mRNA. The kit is compatible with any bacterial species and input RNA prepared from multiple sources, such as total RNA, bacterial mRNA or RNA. aRNA generated by the MessageAmp™ II-Bacteria Kit can be labeled by either inclusion of a labeled nucleotide in the IVT reaction, or by reverse transcription of the aRNA in a reaction containing a labeled nucleotide. The MessageAmp™ II-Bacteria Kit can thus be used to prepare both 33P-labeled sense strand cDNA (for nylon PCR arrays) and biotin- or fluorescent dye–labeled aRNA for glass slide microarrays or Affymetrix GeneChip® arrays.

Accessory products
Bacterial mRNA can be enriched from total RNA using the MICROBExpress™ Bacterial mRNA Enrichment Kit (Cat. No. AM1905). Bacterial RNA can be separated from host cell mixtures with the MICROBEnrich™ Kit (Cat. No. AM1901).

WT Expression Kit, high throughput Invitrogen™

The WT Expression Kit is an RNA amplification kit that is designed to generate amplified sense-strand cDNA ready for fragmentation and labeling using the Affymetrix® GeneChip® WT Terminal Labeling Kit.

Features of the WT Expression Kit:

• Optimized for use with Affymetrix® GeneChip® Human, Mouse, and Rat 1.0 ST Arrays
• Allows RNA samples as small as 50 ng to be analyzed on Affymetrix® GeneChip® Arrays
• Supports a streamlined workflow that does not require a separate rRNA depletion step
• Is ideal for high-sensitivity expression profiling

Eliminates separate rRNA depletion step, preserves transcriptome coverage

The first-generation Affymetrix® amplification method requires depletion of ribosomal RNA (rRNA) from RNA samples for optimal exon-level analysis. Conversely, the Ambion® WT Expression Kit uses a novel reverse transcription (RT) priming method that eliminates the need for a separate rRNA depletion step. The kit includes RT primers designed using a proprietary oligonucleotide matching algorithm that eliminates primer sequences with homology to known ribosomal RNAs. The result is complete and unbiased coverage of the transcriptome, with rRNA amplification levels significantly lower than those of other methods.

Consistent results from less input RNA
With the Ambion® WT Expression Kit, samples as small as 50 ng of total RNA can be analyzed on Affymetrix® GeneChip® Human, Mouse, and Rat Exon and Gene 1.0 ST Arrays. Previous methods required 1 µg of total RNA—often difficult or impossible to obtain from limited sources such as stem cells or small tissue samples. The modest requirement for input RNA permits the analysis of rare samples and provides for more efficient and cost-effective experimentation for most sample types. To demonstrate the performance of the WT Expression Kit, RNA from three sample types (HeLa cells, and Microarray Quality Control (MAQC) A and B samples) was prepared in triplicate using either the Affymetrix® GeneChip® WT cDNA Synthesis and Amplification Kit or the WT Expression Kit, and analyzed on Human Exon 1.0 ST Arrays. Total RNA (1 µg) prepared by the Affymetrix® protocol underwent an rRNA-depletion step while just 50 ng total RNA (20-fold less) was prepared for microarray analysis using the WT Expression Kit. Both sets of samples were handled according to the manufacturers' recommendations. Direct correlation of log2 ratios for MAQC samples A and B was high, (r >0.94) at both the exon and transcript levels. Correlation of log2 ratios with gene expression data obtained using TaqMan® Gene Expression assays for real-time PCR was also high for both kits (see Product Bulletin for data).

Identify more differential expression with less RNA
In addition to its lower input RNA requirement, the WT Expression Kit provides a significant increase in sensitivity. A greater number of probe sets detected above background was obtained at the exon level with the WT Expression Kit as a result of an increased signal-to-noise ratio. A comparison of differential expression analysis results from the two methods indicated that, while differentially expressed genes and exons of MAQC A and MAQC B samples were similar between the two kits (78% and 89%, respectively), significantly more differential expression was observed on arrays hybridized with samples prepared using the WT Expression Kit. 499 more genes and 6,850 more exons were found to be differentially expressed in samples prepared using the Ambion® kit, compared to samples prepared with the Affymetrix® kit.

Note: For high-throughput robotics.

Arcturus™ RiboAmp™ PLUS Kit Applied Biosystems™

The ARCTURUS® RiboAmp® PLUS RNA Amplification Kit permits the researcher to perform molecular analysis on RNA samples that were previously considered too small for microarray analysis. The RiboAmp® PLUS Kit offers unique linear amplification from as little as 1 ng of total RNA. Using a proprietary process for high-efficiency, high-fidelity linear amplification, messenger RNA is amplified up to 1000-fold in one synthesis round and up to 1,000,000-fold in two rounds. The RiboAmp® PLUS Kit produces amplified antisense RNA (aRNA), ready for labeling and microarray hybridization or real-time PCR. The RiboAmp Plus Kit can be used to generate biologically relevant results with cDNA and oligonucleotide array platforms. We also offer RiboAmp® HS PLUS Kits for smaller RNA quantities.

Key product features:
• High fidelity – maintain native expression levels
• Greater sensitivity – detect low-abundance mRNA
• High reliability – deliver reproducible amplified RNA for microarray experiments
• Speedy application – process samples faster
• Enhanced specifity – reveal differences in gene expression between cell types

Maintain Native Expression Levels
The RiboAmp® PLUS RNA Amplification Kit offers unprecedented high-fidelity RNA amplification even from microscopic samples. Comparison of microarray results from total cellular RNA and from aRNA amplified using the RiboAmp® PLUS Kit reveal very high concordance between differentially expressed genes. This demonstrates the RiboAmp® PLUS Kit's ability to deliver the amplification fidelity needed for reliable data analysis from small samples.

Detect Low-abundance mRNA
The RiboAmp® PLUS RNA Amplification Kit enables amplification of mRNA in all abundance classes. Using RT-PCR, low-, medium-, and high-abundance genes are readily detected within the amplified RNA population. Amplification of mRNA in all abundance classes ensures that differential gene expression patterns will be identified.

Deliver Reproducible Amplified RNA for Microarray Experiments
The RiboAmp® PLUS RNA Amplification Kit consistently delivers aRNA ready for labeling and hybridization to expression microarrays with minimal amplification-to-amplification variability. Microarray analysis of independent amplifications from the same source RNA shows excellent correlation.

Process Samples Faster
The RiboAmp® PLUS Kit includes complete reagents and devices for template synthesis, transcription, and nucleic acid purification. Tedious vacuum concentrations are completely eliminated by incorporating optimized chemistries and by using Arcturus' novel, high-recovery MiraCol™ Purification Columns. There are significantly fewer pipetting steps because the sample is eluted directly into reaction tubes and reagents are conveniently pre-dispensed and pre-mixed. In one eight-hour workday, the RiboAmp® PLUS Kit enables amplification, labeling, and microarray hybridization set up. With a convenient overnight in vitro transcription, a second round of amplification can be completed by the following morning.

Reveal Differences in Gene Expression between Cell Types
Accurate gene expression analysis requires the analysis of specific cell types without interference from surrounding cells. Starting with these pure cell populations often means working with small samples. Special technologies are needed to overcome the challenges of handling these precious samples. The combination of LCM, RNA amplification, and microarray analysis reveals differential gene expression between cell types.

Integrated Systems for Microgenomics
The Arcturus® Complete System for Microgenomics® offers an integrated solution for preparing small quantities of RNA for gene expression analysis. The System features the ArcturusXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations, the PicoPure® RNA Isolation Kit to extract and purify RNA, and the RiboAmp® PLUS Kit for linear amplification of RNA. Arcturus' Systems for Microgenomics enable accurate and sensitive microarray assays that reveal molecular otherwise obscured in whole tissue samples or cell mixtures.

Arcturus™ RiboAmp™ PLUS Kit, Cy3 labeling Applied Biosystems™

The ARCTURUS® RiboAmp® PLUS RNA Amplification Kit enables the researcher to perform molecular analysis on RNA samples that previously would have been considered too small for microarray analysis. The RiboAmp® PLUS Kit offers unique linear amplification from as little as 1 ng of total RNA. Using a proprietary process for high-efficiency, high-fidelity linear amplification, messenger RNA is amplified up to 1000-fold in one synthesis round and up to 1,000,000-fold in two rounds. The RiboAmp® PLUS Kit produces amplified antisense RNA (aRNA), ready for labeling and microarray hybridization or real-time PCR. The RiboAmp® PLUS Kit can be used to generate biologically relevant results with cDNA and oligonucleotide array platforms.

Key product features:
• Robust, high quality kit for microarray labeling and Gene Expression profiling
• Flexibility in Assay Format

Microarray Labeling, Gene Expression Profiling
The RiboAmp® PLUS RNA Amplification Kit may be configured to include Turbo Labeling™ for use in microarray experiments. Turbo Labeling™ Kits offer a proprietary, non-enzymatic technology optimized for the labeling of unmodified, amplified RNA (aRNA) for gene expression profiling. The unmodified aRNA is labeled post-amplification, thereby avoiding the need to incorporate modified nucleotides during RNA amplification. The use of unmodified nucleotides in the amplification process results in unmodified aRNA with higher yields and longer aRNA fragments, thus providing better representation of the mRNA transcript for downstream analysis (Figure 1).

Flexibility in Assay Format
Turbo Labeling Kits include reagents to support labeling of 12 samples using either Cy™3⁄Cy™5 dyes or Biotin for hybridization to cDNA or oligonucleotide arrays. Figure 1 demonstrates the process for these kits and how to choose the kit for your research. Both provide reproducibile results that allow you to detect more genes (Figures 2 & 3).

Integrated Systems for Microgenomics
Arcturus' complete Systems for Microgenomics® offers an integrated solution for preparing small quantities of RNA for gene expression analysis. The System features the ArcturusXT™ Laser Capture Microdissection (LCM) Instrument to procure pure cell populations, the PicoPure® RNA Isolation Kit to extract and purify RNA, the RiboAmp® PLUS Kit for linear amplification of RNA and the Turbo Labeling™ Kit to prepare the aRNA for microarray analysis. ARCTURUS®' Systems for Microgenomics enable accurate and sensitive microarray assays that reveal molecular signatures otherwise obscured in whole tissue samples or cell mixtures.

For Research Use Only. Not for use in diagnostics procedures.

Illumina™ TotalPrep™-96 RNA Amplification Kit Invitrogen™

The TotalPrep™-96 RNA Amplification Kit is a complete system for generating biotinylated, amplified RNA for hybridization with Illumina Sentrix® arrays. The kit includes sufficient reagents for 96 reactions. Features of the TotalPrep™-96 RNA Amplification Kit:

• Perform amplification from just 50 ng of input RNA
• Developed for expression profiling using the Sentrix® BeadChips
• Bio-16-UTP now included in optimized NTP mix, reducing set-up time and maximizing labeling

Using the TotalPrep™-96 RNA Amplification Kit
The TotalPrep™-96 RNA Amplification Kit incorporates extensive improvements to the in vitro transcription reaction (IVT) over the previous TotalPrep™ Kit. The IVT reaction has been optimized for the most effective biotin labeling. An NTP mix containing biotinylated UTP has been added to the kit, allowing ease of use and optimal labeling. These modifications to the kit have significantly increased sensitivity, providing increased Percent Present calls and single-round amplification with just 50 ng of input RNA. The kit is based on the RNA amplification protocol developed in the laboratory of James Eberwine (Van Gelder et al. 1990). The procedure consists of reverse transcription with an oligo(dT) primer bearing a T7 promoter using ArrayScript™, a reverse transcriptase (RT) engineered to produce higher yields of first strand cDNA than wild type enzymes. ArrayScript™ catalyzes the synthesis of virtually full-length cDNA, which is the best way to ensure production of reproducible microarray samples. The cDNA then undergoes second strand synthesis and magnetic-based cleanup to become a template for in vitro transcription with T7 RNA Polymerase. To maximize cRNA yield, MEGAscript® in vitro transcription (IVT) technology, along with biotin UTP (provided in the kit), is used to generate hundreds to thousands of biotinylated, antisense RNA copies of each mRNA in a sample.

Accessory product
A 96-well magnetic-ring stand is required to use this kit. The 96-well Magnetic-Ring Stand (Cat. No. AM10050) is recommended and designed for paramagnetic bead precipitation from standard 96-well U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors.

MessageAmp™ III RNA Amplification Kit Invitrogen™

This Ambion® aRNA amplification kit has extensive improvements over existing methods. It has fewer components, pipetting steps, and manipulations in the workflow which shorten hands-on time, and reduce operator-dependant errors and variability. The kit uses a glass fiber, filter-based aRNA purification method. The kit includes sufficient reagents for 10 reactions.

Simplified Workflow
• All reactions occur in a single tube
• Master Mix formulations reduce pipetting steps and hands-on time
• Shortened processing time due to optimized incubations and condensed procedure
• Less hands-on time means less opportunity to introduce errors

Improved Reaction Efficiency
• Start with as little as 20 ng total RNA and produce enough aRNA for GeneChip® analysis
• Start with 100 ng of total RNA (with a 4 hour IVT) and hybridize to a GeneChip® array in a single day

DNA microarrays are used to evaluate the expression levels of thousands of genes in a single experiment. High-density arrays are the method of choice for assessing gene expression on a genome-wide scale. One of the challenges associated with microarray technology, however, is that adequate amounts of labeled antisense RNA (known as cRNA or aRNA) are required. T7 linear amplification, commonly referred to as the Eberwine method, overcomes this challenge by simultaneously amplifying and labeling the RNA. This method is the basis of the MessageAmp™ III RNA Amplification Kit.

Workflow Improvements
The kit includes a set of master mixes, reducing the number of components, pipetting steps, and manipulations. Steps such as initial primer annealing and cDNA purification have been eliminated. All steps of the biotin labeling/amplification process take place in a single reaction tube. This simplified workflow significantly shortens hands-on time, reducing operator-dependant errors and variability.

Improved Reaction Efficiency
The MessageAmp™ Premier Kit allows a researcher to perform expression profiling with as little as 20 ng of input RNA. The kit requires as little as 100 ng of total RNA, producing enough labeled aRNA to hybridize to a GeneChip® array in a single day. Representative yields from the MessageAmp™ Premier Kit using HeLa total RNA and RNA from multiple tissue types are shown in the data.

MessageAmp™ II-Biotin Enhanced aRNA Amplification Kit Invitrogen™

The Ambion® MessageAmp™ II-Biotin Enhanced aRNA Amplification Kit is for the amplification of as little as 50 pg of input RNA for array analysis. The included NTP mix contains biotinylated UTP. The kit includes sufficient reagents for 20 single-round aRNA amplifications.

• Detect more genes due to increased percent present calls
• Obtain sufficient amounts of amplified RNA from a single round of amplification with only 50 ng of input RNA
• Biotinylated nucleotide is included in the optimized NTP mix, reducing setup time and maximizing labeling

Expression profiling using gene arrays has become the most widely used method for assessing gene expression on a genome-wide scale. The primary limitation of this technology is the requirement for large amounts of RNA for array hybridization. This hurdle is usually overcome by using T7 RNA Polymerase–driven transcription of cDNA (obtained from total RNA samples) to amplify sufficient amounts of cRNA. This amplification method is the basis of the MessageAmp™ II-Biotin Enhanced aRNA Amplification Kit.

Improvements for Biotin Labeling
The IVT reaction step of the MessageAmp™ II-Biotin Enhanced Kit procedure has been optimized for effective biotin labeling. An NTP mix containing biotinylated UTP has been added to the kit for ease of use. Other optimizations have increased sensitivity and increased yields so that just 50 ng of input RNA is sufficient to produce enough aRNA for array analysis after just a single round of amplification. These kits do not include enough reagents for two rounds of amplification. If your experiments require two rounds of amplification, you will need to purchase the MessageAmp™ II aRNA Amplification Kit (see below) in addition to the MessageAmp™ II-Biotin Kit.

Accessory Product
The MessageAmp™ II aRNA Amplification Kit (SKU#AM1751)is available should additional reagents be needed for two rounds of aRNA amplification.
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