Turbo Labeling Kits

aRNA Labeling For Gene Expression Profiling

Turbo Labeling™ Kits offer a proprietary, non-enzymatic technology optimized for the labeling of unmodified, amplified RNA (aRNA) for gene expression profiling. The unmodified aRNA is labeled post-amplification, thereby avoiding the need to incorporate modified nucleotides during RNA amplification. The use of unmodified nucleotides in the amplification process results in unmodified aRNA with higher yields and longer aRNA fragments, thus providing better representation of the mRNA transcript for downstream analysis.


Turbo Labeling Kits include reagents to support labeling of 12 samples using either Cy™3*/Cy™5 dyes or Biotin for hybridization to cDNA or oligonucleotide arrays.
  • Simple aRNA labeling protocol takes less than 30 minutes
  • Improved efficiency
  • Unlabeled aRNA can be preserved for downstream validation
  • Platform-independent protocol
  • Higher yields

Turbo Labeling of unmodified aRNA provides flexibility in assay format choice


Schematic representation of the Turbo Labeling process for microarray hybridization


Uniform Labeling

Reproducibility of Cy3 and Cy5 labeling. 10 ng of Universal Human Reference total RNA was amplified in duplicate using the RiboAmp RNA Amplification Kit and 15 μg aRNA from each replicate amplification was labeled with Turbo Labeling Cy3 & Cy5 Kits and hybridized onto an oligonucleotide array. Spots whose median intensity were ≥ 2× background intensity and within-spot CV of intensities ≤ 20% were plotted on a scatter-plot and Pearson correlation coefficient (r) was determined. A: Replicate amplifications and labeling with Cy3. B: Replicate amplifications and labeling with Cy5. C: Comparison between Cy3 and Cy5 labeling. Comparison of normalized gene intensities showed excellent reproducibility across all genes (r = 0.97-0.98), suggesting comparable labeling efficiencies for Cy3 and Cy5 dyes and highly reproducible array data.          

Detect More Genes
                     
Percent present (%P) calls from Turbo Labeling Biotin Kit compared to an alternate Biotin labeling kit from Company A. 10 ng and 2 μg of Universal Human Reference total RNA were amplified in duplicate using the RiboAmp® RNA Amplification System, and biotinylated aRNA were prepared using either the Turbo Labeling Biotin Kit or Company A’s Biotin labeling kit. 12.8 μg of each labeled aRNA was hybridized onto the GeneChip Human Genome U133 Plus 2.0 Array. Turbo Labeling generated higher %P calls at both RNA input levels.          
                     
Stringent Reproducibility
                     
Correlation of signal intensity obtained from two GeneChip® Arrays comparing the Turbo Labeling Biotin Kit and a commercially available Biotin labeling kit. Two separate samples (2 μg) of Universal Human Reference total RNA were amplified with RiboAmp RNA Amplification kit and labeled using either the Turbo Labeling Biotin Kit or Company A’s Biotin labeling kit. Labeled samples were hybridized overnight to GeneChip
Human Genome U133 plus 2.0 Array. Highly reproducible gene intensity values were obtained between genes called “present” in both methods (Pearson correlation, r = 0.96).

*This product or portions thereof is manufactured under license from Carnegie Mellon University under U.S. Patent Numbers [5,268,486 ] and [4,981,977] and related patents. Cy and CyDye are trademarks of GE Healthcare Limited.

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