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MicroRNA Profiling by Array Analysis Reveals Critical BioMarkers

ists have developed a microarray analysis procedure that interrogates only the mature and functionally active miRNAs (See Novel MicroRNA Array Technology for Sensitive miRNA Profiling). The procedure comprises four steps and relies on many of the processes and instrumentation that are used for standard mRNA microarray analysis:

Step 1 - Total RNA that includes the small RNA fraction is isolated from a tissue or cell sample using the mirVana miRNA Isolation Kit (Ambion).

Step 2 - The RNA in the sample is fractionated using a specialized electrophoresis unit and the miRNA fraction is recovered.

Step 3 - The miRNA sample is tailed by poly(A) polymerase. The tail includes an amine-modified nucleotide that reacts with activated Cy3 or Cy5 to fluorescently label the miRNAs. The mirVana miRNA Labeling Kit (Ambion) makes the tailing and subsequent purification steps simple, and more importantly, reproducible.

Step 4 - The fluorescent miRNAs are hybridized to a glass slide upon which specially designed miRNA-specific probes have been arrayed. miRNA probes for printing glass miRNA Arrays are available from Ambion as the mirVana miRNA Probe Set (Ambion). The resulting miRNA arrays are then processed using standard array scanners.


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