MessageAmp™ III RNA Amplification Kit

Catalog number: AM1793

Ambion™  Related applications: Microarray Analysis

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Description

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 30 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 the 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.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 30 reactions
Labeling Method: Direct Labeling
Includes Label or Dye: Yes
Reverse Transcriptase: ArrayScript™ Reverse Transcriptase
Sample Type (Specific): Total RNA, Poly(A+) RNA

Contents & storage

The MessageAmp™ III aRNA Amplification Kit includes First-Strand Enzyme Mix, First-Strand Buffer Mix, Second-Strand Enzyme Mix, Second-Strand Buffer Mix, T7 Enzyme Mix, T7 Biotin IVT Mix, Control RNA (1 mg⁄mL HeLa total RNA) which are all stored at –20°C. Wash Buffer is stored at +4°C and aRNA Binding Buffer, Filter Cartridges, Collection Tubes, 8-Strip PCR Tubes & Caps (0.2 mL), 5X Array Fragmentation Buffer, and Nuclease-free Water are stored at room temperature.

Documents

Manuals & protocols