A Systematic Comparison of Purification and Normalization Protocols for Quantitative MicroRNA Expressional Profiling in Insulin-Producing Cells
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A Systematic Comparison of Purification and Normalization Protocols for Quantitative MicroRNA Expressional Profiling in Insulin-Producing Cells. / Vestergaard, Anna Lindeløv; Blankestijn, Maaike; Stahl, Jonathan Lucien; Pallesen, Emil Marek Heymans; Bang-Berthelsen, Claus Heiner; Pociot, Flemming; Novotny, Guy Wayne; Lundh, Morten; Mandrup-Poulsen, Thomas.
In: International Journal of Molecular Sciences (Online), Vol. 17, No. 6, 896, 2016.Research output: Contribution to journal › Comment/debate › Research › peer-review
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TY - JOUR
T1 - A Systematic Comparison of Purification and Normalization Protocols for Quantitative MicroRNA Expressional Profiling in Insulin-Producing Cells
AU - Vestergaard, Anna Lindeløv
AU - Blankestijn, Maaike
AU - Stahl, Jonathan Lucien
AU - Pallesen, Emil Marek Heymans
AU - Bang-Berthelsen, Claus Heiner
AU - Pociot, Flemming
AU - Novotny, Guy Wayne
AU - Lundh, Morten
AU - Mandrup-Poulsen, Thomas
PY - 2016
Y1 - 2016
N2 - As microRNAs (miRs) are gaining increasing attention as key regulators of cellular processes, expressional quantification is widely applied. However, in the processing of relatively quantified data, the importance of testing the stability of several reference mRNAs and/or miRs and choosing among these for normalization is often overlooked, potentially leading to biased results. Here, we have optimized the purification of miR-enriched total RNA from pancreatic insulin-producing INS-1 cells. Additionally, we optimized and analyzed miR expression by a qPCR-based microarray and by specific qPCR and tested the stability of candidate reference mRNAs and miRs. Hence, this study gives a widely applicable example on how to easily and systematically test and decide how to normalize miR quantification. We suggest that caution in the interpretation of miR quantification studies that do not comprise stability analysis should be exerted.
AB - As microRNAs (miRs) are gaining increasing attention as key regulators of cellular processes, expressional quantification is widely applied. However, in the processing of relatively quantified data, the importance of testing the stability of several reference mRNAs and/or miRs and choosing among these for normalization is often overlooked, potentially leading to biased results. Here, we have optimized the purification of miR-enriched total RNA from pancreatic insulin-producing INS-1 cells. Additionally, we optimized and analyzed miR expression by a qPCR-based microarray and by specific qPCR and tested the stability of candidate reference mRNAs and miRs. Hence, this study gives a widely applicable example on how to easily and systematically test and decide how to normalize miR quantification. We suggest that caution in the interpretation of miR quantification studies that do not comprise stability analysis should be exerted.
U2 - 10.3390/ijms17060896
DO - 10.3390/ijms17060896
M3 - Comment/debate
C2 - 27338345
VL - 17
JO - International Journal of Molecular Sciences (Online)
JF - International Journal of Molecular Sciences (Online)
SN - 1661-6596
IS - 6
M1 - 896
ER -
ID: 165754813