Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Data on the effect of heat and other technical variables on the detection of microRNAs in human serum.

Author(s): Camacho L, Porter-Gill P, Silva CS

Publication: Data Brief, 2019, Vol. 24, Page 103750

PubMed ID: 30976632 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to investigate the effects of heating serum specimens prior to extraction, extraction method used, and real-time PCR array type on the number of miRNAs detected and the distribution of CT values. Four commercially-obtained frozen human serum specimens were divided into two aliquots each. One aliquot was heated at 60˚C for 120 min and the other was not heated. Heated and not heated aliquots were divided and miRNA was extracted using both the miRCURY RNA Isolation Kit for Biofluids and the SeraMir Exosome RNA Purification Kit. miRNA was reverse-transcribed using the TaqMan Micro-RNA Reverse Transcription Kit and quantified by the real-time PCR-based TaqMan Low Density Human microRNA Array A or reverse-transcribed using the miRCURY LNA Universal cDNA Synthesis Kit II and quantified using the Exiqon MicroRNA Ready-to-use PCR Human Panel I V4.M Array.

   Summary of Findings:

   CT values tended to be lower for the TaqMan Low Density Human microRNA Array A than the Exiqon MicroRNA Ready-to-use PCR Human Panel I V4.M. The authors attribute this difference to the inclusion of a preamplification step in the Applied Biosystems protocol. Extraction method had little effect on the number of miRNAs detected or the distribution of the CT values when the RNA was not heated before extraction. However, heating serum prior to extraction resulted in the detection of fewer miRNAs using either array when extracted using the SeraMir Exosome RNA Purification Kit but had no effect when extraction was with the miRCURY RNA Isolation Kit.

   Biospecimens

   • Bodily Fluid - Serum

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR
   RNA	Low density array

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	miRCURY RNA Isolation Kit for Biofluids SeraMir Exosome RNA Purification Kit
   Real-time qRT-PCR Specific	Technology platform	TaqMan Low Density Human microRNA Array A Exiqon MicroRNA Ready-to-use PCR Human Panel I V4.M Array
   Storage	Storage duration	0 h 2 h at 60˚C

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