Ambion™ ArrayControl™ RNA Spikes consist of 8 individual RNA transcripts ranging in size from 750 to 2,000 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. Each transcript is supplied in one tube containing 1 μg.
• 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 When hybridized to complementary spots on a gene array, ArrayControl™ 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, 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 have been 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. Their manufacturing and quantitation are consistent from lot-to-lot, so that signals can be compared between experiments and to assess array reproducibility. To obtain RNA Spike sequence information for the preparation of complementary control spots on your arrays, please contact Technical Support.