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Figure 2 | BMC Genomics

Figure 2

From: An optimized protocol for microarray validation by quantitative PCR using amplified amino allyl labeled RNA

Figure 2

Amplification and amino allyl labeling decrease qPCR efficiency and sensitivity. One μg RNA and 100 ng AA-aRNA from human universal reference were reverse transcribed with SuperScript II. (A) Ten-fold dilutions of resulting cDNAs were subjected to qPCR using primer pairs specific for VEGFB. The amplification chart shows higher Cq values for AA-aRNA compared with RNA. (B) cDNAs obtained from reverse transcription of RNA and AA-aRNA were serially diluted and qPCR was performed to evaluate qPCR efficiency using VEGFB-specific primers. qPCR efficiency calculated from standard curves (E = [10-1/slope]-1) is down-regulated by amplification and amino allyl labeling.

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BMC Genomics

ISSN: 1471-2164

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