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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 magnetic bead–based aRNA purification method and 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 • Magnetic bead-based purification gives high yields with more consistent results
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™ Premier 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.
Advantages of Magnetic Bead–based aRNA Purification The MessageAmp™ Premier Kit employs a magnetic bead–based method for aRNA clean-up. Advantages to this method include: • Higher yields (up to 25%) than glass fiber filter purification • More consistent results • No filters to clog • Faster workflow • Easier to handle using manual pipetting • More amenable to high-throughput processing
Accessory Product The Magnetic Stand-96 (SKU#AM10027)is designed for paramagnetic bead precipitation from standard 96-well U-bottom microplates and 0.2 mL PCR plates with no additional accessories. The stand has a clear polyacryl base in a standard SBS 96-well microplate footprint. The use of 24 extremely strong permanent magnets guarantees easy and fast (as little as 30 seconds) magnetic separation. It sits flat on a lab bench, as well as in a microplate adapter on robotic workstations.
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