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Use of Transcriptor Reverse Transcriptase,,,in Microarray Analysis

RNA was used in all experiments, it was possible to test an enzymes ability to generate reproducible results; microarrays using Transcriptor Reverse Transcriptase and product Y were replicated four times each. Ratio (Cy5/Cy3) data from each of the four replicates plotted against the mean show similar results, where the Transcriptor Reverse Transcriptase replicates show generally better correlation to the mean than product Y (Figure 3). This appears to be due to fewer outlier spots that depart substantially from the trend lines in the four Transcriptor Reverse Transcriptase replicates.



The replicate data were processed in GeneSpring to detect particular spots that are consistently biased towards greater signal in either Transcriptor Reverse Transcriptase or product Y. Inspection of a few cDNA probes showed that the greatest signal bias towards Transcriptor Reverse Transcriptase appear to be in those probes that are substantially truncated, and those with a greater signal bias towards product Y are truncated much closer to the 5′ end. Although this anecdotal observation could be interpreted in several ways, it is at least consistent with the notion that the high processivity of Transcriptor Reverse Transcriptase may produce a better labelled target and more robust data in cases where the spot/probe cDNA is substantially truncated.

Conclusion

The histogram of signal intensity distributions show that Transcriptor Reverse Transcriptase can be used for high efficient target labelling. Since many investigators eliminate data with signal intensities below a particular threshold (e.g.,1,0002,000), an increased specific activity of the labelled target may be critical for analysing genes with low expression.

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