Circulating miRNA Profiling in Blood

Expression profiling of circulating miRNAs was performed on RNA extracted from 20 blood specimens using TaqMan human miRNA microarrays as instructed by the manufacturer (TaqMan Low Density Array Human microRNA Card A, Life Technologies, Foster City, CA, USA). Megaplex primer pools were used to reverse transcribe RNA samples (700 ng) which were then PCR amplified in 2 µL volumes on 384 well pre-configured microfluidic cards. Each card contained TaqMan probes for 377 miRNAs plus 3 pre-defined ECs (U6 in quadruplicate, RNU44 and RNU48) and a negative control (ath-miR-159a).

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McDermott A.M., Kerin M.J, & Miller N. (2013). Identification and Validation of miRNAs as Endogenous Controls for RQ-PCR in Blood Specimens for Breast Cancer Studies. PLoS ONE, 8(12), e83718.

Publication 2013

Blood Circulating mirnas Human Microarrays Mirnas Primer

Corresponding Organization : Ollscoil na Gaillimhe – University of Galway

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Protocol cited in 10 other protocols

Variable analysis

independent variables

Expression profiling of circulating miRNAs

dependent variables

Measured outcomes of miRNA expression

control variables

RNA extracted from 20 blood specimens
TaqMan human miRNA microarrays
Megaplex primer pools used for reverse transcription
PCR amplification in 2 µL volumes on 384 well pre-configured microfluidic cards
TaqMan probes for 377 miRNAs
3 pre-defined ECs (U6 in quadruplicate, RNU44 and RNU48)
Negative control (ath-miR-159a)

controls

Positive controls: U6 in quadruplicate, RNU44 and RNU48
Negative control: ath-miR-159a

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