• For the normalization and control of microarray experiments
  
  
• For array printing and protocol assessment/optimization
  
  
• Monitor reverse transcription and cDNA labeling efficiency, hybridization stringency, and background
  
  
• Normalize Cy™3 and Cy™5 signals
  
  

Gene array analysis is a powerful technique for elucidating the expression of thousands of genes in a single experiment. However, gene array analysis is a multi-step procedure with many opportunities for sample, procedural, and user variations. Gene array data is most valid when compared to control sequences that have known concentrations and are mixed at known ratios.

ArrayControl™ RNA Spikes
ArrayControl RNA Spikes consist of 8 individual RNA transcripts ranging in size from 750 to 2000 bases with a 30-base 3' poly(A) tail. RNA Spikes are added to the RNA sample and used as template for preparing labeled cDNA. When hybridized to complementary spots on a gene array, RNA Spikes can be used to monitor the efficiency of reverse transcription and aRNA amplification, labeling, and hybridization. They can also be used to assess the precision, accuracy and consistency of array printing and protocol sensitivity. In addition, ArrayControl RNA Spikes are an invaluable tool for protocol optimization and troubleshooting. They can be used for normalization of Cy™3 and Cy™5 signals, determination of dynamic range, and slide orientation. Individual RNA Spikes can also be omitted to serve as a negative control for background calculations.

The RNA Spike sequences are carefully chosen to lack homology to mammalian sequences that might otherwise lead to cross hybridization to gene specific spots. Each RNA Spike is purified, precisely quantitated by spectrophotometry and RiboGreen® staining, and tested for stability under highly stringent conditions. They are also tested for cDNA labeling efficiency using both direct and indirect methods. Because manufacturing and quantitation of ArrayControl RNA Spikes are consistent from lot-to-lot, signals can be compared to assess array printing reproducibility and to compare data between experiments.

To obtain sequence information for the RNA Spikes for the preparation of complementary control spots on your arrays, please contact Ambion Technical Support.

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