Identification of miR-16 as an endogenous reference gene for the normalization of urinary exosomal miRNA expression data from CKD patients

Abstract BACKGROUND Different technologies, such as quantitative real-time PCR or microarrays, have been developed to measure microRNA (miRNA) expression levels. Quantification of miRNA transcripts implicates data normalization using endogenous and exogenous reference genes for data correction. However, there is no consensus about an optimal normalization strategy. The choice of a reference gene remains problematic and can have a serious impact on the actual available transcript levels and, consequently, on the biological interpretation of data. CONTENT In this review article we discuss the reliability of the use of small RNAs, commonly reported in the literature as miRNA expression normalizers, and compare different strategies used for data normalization. SUMMARY A workflow strategy is proposed for normalization of miRNA expression data in an attempt to provide a basis for the establishment of a global standard procedure that will allow comparison across studies.

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Decision tree‑based classifiers for lung cancer diagnosis and subtyping using TCGA miRNA expression data

Oncology Letters ◽

10.3892/ol.2019.10462 ◽

2019 ◽

Cited By ~ 3

Author(s):

Masih Sherafatian ◽

Fateme Arjmand

Keyword(s):

Lung Cancer ◽

Decision Tree ◽

Cancer Diagnosis ◽

Mirna Expression ◽

Expression Data ◽

Lung Cancer Diagnosis ◽

Mirna Expression Data

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Identification of a chicken ( Gallus gallus ) endogenous reference gene ( Actb ) and its application in meat adulteration

Food Chemistry ◽

10.1016/j.foodchem.2017.05.038 ◽

2017 ◽

Vol 234 ◽

pp. 472-478 ◽

Cited By ~ 14

Author(s):

Wenjin Xiang ◽

Ying Shang ◽

Qin Wang ◽

Yuancong Xu ◽

Pengyu Zhu ◽

...

Keyword(s):

Reference Gene ◽

Gallus Gallus ◽

Endogenous Reference Gene ◽

Endogenous Reference

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Validation of Artemisia Genus-Specific Gene, GEL4, for Use as an Endogenous Reference Gene in Qualitative PCR Detection of Putative Transgenic Artemisia annua

Journal of AOAC International ◽

10.5740/jaoacint.11-358 ◽

2013 ◽

Vol 96 (3) ◽

pp. 587-592

Author(s):

Hua Liu ◽

Lingxi Jiang ◽

Furong Tan ◽

Xinghai Zhao ◽

Heyan Lin ◽

...

Keyword(s):

Reference Gene ◽

Artemisia Annua ◽

Intraspecific Variability ◽

Specific Gene ◽

Endogenous Reference Gene ◽

Medical Plant ◽

Endogenous Reference ◽

Pcr Method ◽

Artemisinin Content ◽

Qualitative Pcr

Abstract Artemisia annua L. is a popular medical plant used against malaria. Transgenic A. annua plants have been developed to increase the artemisinin content using genetic engineering. To develop a reliable PCR method of detecting transgenic A. annua, one Artemisia specific gene, GEL4, was selected and validated as suitable. Qualitative PCR methods were assayed with 16 different A. annua ecotypes and four Artemisia plants, and identical amplified products were obtained. No amplified products were observed when genomic DNA samples were extracted from 15 different plant species. An LOD of 0.05 ng of A. annua DNA was also determined. Furthermore, the results of a Southern blot method showed that the GEL4 gene was in three copies of the six A. annua ecotypes tested. To determine whether there was intraspecific variability in the GEL4 region between different A. annua ecotypes, the GEL4 region corresponding to the 10 A. annua ecotypes was cloned and sequenced. Alignment of the obtained sequences presented a highly conserved amplification product region in 10 A. annua ecotypes. Therefore, it is concluded that the GEL4 gene can be used as an endogenous reference gene of A. annua, and that the qualitative PCR system was reliable for the detection of transgenic A. annua.

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