The ARCTURUS™ RiboAmp™ PLUS RNA Amplification Kit permits the researcher to perform molecular analysis on RNA samples that were previouslyRead more
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Catalog Number
Quantity
KIT0501
24 Samples
Catalog number KIT0501
Price (MXN)
-
Quantity:
24 Samples
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.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
For Use With (Application)Microarray Analysis, Real-Time Quantitative PCR (qPCR)
For Use With (Equipment)ArcturusXT™ LCM Instrument
IncludesAmplification Purification Kit (Purification Reagents and Columns), cDNA Synthesis Kit, In vitro Transcription (IVT) Kit
Workflow StepReverse Transcription, In Vitro Transcription, RNA Amplification
Unit SizeEach
Contents & Storage
cDNA synthesis kit, in vitro transcription (IVT) kit, amplification purification kit (purification reagents and columns). Variable storage conditions
Frequently asked questions (FAQs)
What is the typical size range of amplified RNA?
A single round of amplification yields product sizes ranging from 200 bases to 6 kb. The majority of these products are approximately 1.5 kb in length. A second round of amplification will result in shorter products. We recommend using an Agilent 2100 bioanalyzer to visualize these products. Amplification products can be visualized by agarose gel electrophoresis; they will migrate as a smear. Although this data is still useful, it is less informative than bioanalyzer analysis.
Glass microarray analysis experiments typically require 5-20 µg of total RNA per slide for sample labeling and hybridization. Thus, microarray-based gene expression analysis of very small samples [laser capture microdissection (LCM), tissue biopsies, or other clinical samples] is difficult due to the very low amounts of total RNA recovered from the samples. Linear amplification of RNA from small samples produces sufficient quantities of RNA for sample labeling and hybridization. Since the amplification technique is highly reproducible and maintains representation of the gene expression in the original sample, it is recommended for probe synthesis by most manufacturers of commercially available microarrays.
RNA amplification using the Van Gelder and Eberwine technique (Van Gelder 1990) utilizes an oligo(dT) primer containing the T7 RNA polymerase promoter for synthesis of first strand cDNA. The poly(A) tail at the end of mRNA sequences serves as the substrate for the binding of these primers. Since mRNA typically constitutes only 1-5% of the total RNA in the cell, only this fraction of the total RNA is amplified. The tissue type, its developmental state, and its health all influence the actual proportion of mRNA in a total RNA sample. Total RNA from brain, testes, and embryonic tissues may contain up to 4% mRNA, while RNA from many other tissues will have only 1% or less mRNA. The RNA isolation method can also influence mRNA content. The generally accepted average value for mRNA content is about 2% of a total RNA sample. When 1 µg of total RNA, 2% or 20 ng of which is mRNA, is amplified 1000-fold, yields of 20 µg aRNA (or cRNA) should be expected. You may observe higher fold amplification when starting with lower amounts of total RNA. This is because, in an in vitro transcription (IVT) reaction, a finite amount of RNA can be synthesized with the fixed amount of NTPs. When starting with less RNA, NTPs do not become limiting until the RNA is amplified beyond the typical 1000-2000 fold amplification levels seen with higher amounts of input RNA.