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Bioanalysis. 2013 Jul;5(13):1623-33. doi: 10.4155/bio.13.125.

Evaluation of a high-throughput, microfluidics platform for performing TaqMan™ qPCR using formalin-fixed paraffin-embedded tumors.

Author information

  • 1Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA.

Abstract

BACKGROUND:

Compared with the standard qPCR, nanoliter-scale qPCR can use smaller quantities of RNA and increase throughput. The TaqMan™ OpenArray® NT Cycler System was assessed for use with degraded RNA from formalin-fixed paraffin-embedded (FFPE) tumors.

RESULTS:

Expression of candidate prognostic genes was quantified using the OpenArray platform and matching fresh frozen and FFPE patient renal cell carcinomas. Reverse transcription, with gene-specific reverse transcription and preamplification, optimized the percentage of detectable transcripts. When using high quality RNA from fresh frozen tumors, the OpenArray platform identified 30 prognostic genes. However, when using RNA from FFPE tumors, only 13 prognostic genes were identified, but this increased to 33 with addition of preamplification.

CONCLUSION:

The OpenArray platform can be optimized to quantify gene expressions from FFPE tumors.

PMID:
23822126
[PubMed - indexed for MEDLINE]
PMCID:
PMC3816109
Free PMC Article

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