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Figure 3 | BMC Biotechnology

Figure 3

From: Improved RNA quality and TaqMan®Pre-amplification method (PreAmp) to enhance expression analysis from formalin fixed paraffin embedded (FFPE) materials

Figure 3

TaqMan® gene expression pattern using UPMM, GEMM and PreAmp. (Str = Stratagene protocol; Am = Ambion protocol; + = modified protocols with incubation in Proteinase K buffer at 70°C for 20 minutes; and SN = snap frozen cells. Not all assays produced products, e.g. HLA_A in UPMM.) CTs were higher using FFPE extracts compared to snap frozen counterparts when the amount of input RNA was identical. Modification to Stratagene and Ambion protocols produced decreased CTs. TaqMan® with UPMM generated higher CTs than GEMM and PreAmp when using longer amplicon lengths. This was particularly evident when comparing two GAPDH assays – one 122 bp and the other 67 bp. A mean difference of 17 cycles between small and large assays using UPMM and a mean difference of 4 cycles between small and large assays using GEMM was observed.

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BMC Biotechnology

ISSN: 1472-6750

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