Detectability of cir detectability of circulating micr culating microRNAs in micr As in microRNA extracts with acts with low purity and yield using quantitative real-time polymerase chain reaction: Supporting evidence

Ahmad, Azmir and Kaderi, Mohd Arifin and Tumian, Afidalina and Sivanesan, Vijaya Mohan and Abdullah, Kahairi and Leman, Wan Ishlah and Mohamad, Irfan and Wan Zainon, Wan Mohd. Nazri and Mohd. Shiyuti, Muhammad Izani and Mohamed Awang, Kamariah and Rosla, Luqman and Paul, Mark and Syed Yussof, Sharifah Nor Ezura and Ramli, Rosdi (2020) Detectability of cir detectability of circulating micr culating microRNAs in micr As in microRNA extracts with acts with low purity and yield using quantitative real-time polymerase chain reaction: Supporting evidence. Makara Journal of Health Research, 24 (3). pp. 147-156. ISSN 2356-3664 E-ISSN 2356-3656

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Abstract

Background: Circulating microRNAs (miRNAs) are a group of noncoding RNAs with promising potential as minimal invasive biomarkers for noncommunicable diseases. However, challenges exist in the preparation of these miRNAs from peripheral blood samples for quantification purposes. The low quality of miRNA extracts presents an obstacle. Acknowledging the superior performance of quantitative real-time polymerase chain reaction (qPCR) as gold standard for gene expression analysis, we conducted this study to observe the capabilities of qPCR using the Taqman® protocol in amplifying circulating miRNAs from miRNA extracts with low purity and yield. Methods: miRNAs were extracted from thirty-six plasma samples that were obtained from public subjects. Four selected miRNAs were quantified using the Taqman® protocol in an integrated fluidic circuit chip that was optimized from a previous study. The amplification graph and Cq values were obtained to observe any abnormal amplification signs and expression levels, respectively. Results: The qualitative observation of the amplification of the four miRNAs showed no sign of abnormality, thereby indicating the successful amplification of the miRNAs without enzymatic inhibition. Furthermore, the miRNAs were quantified in high expression levels. Conclusion: The circulating miRNA extracts with low purity and yield were practical for the study of circulating miRNA expression based on the Taqman® protocol as the method of detection.

Item Type:	Article (Journal)
Uncontrolled Keywords:	amplification, circulating, isolation and purification, microRNA, quantitative real-time polymerase chain reaction
Subjects:	R Medicine > R Medicine (General)
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Allied Health Sciences Kulliyyah of Allied Health Sciences > Department of Biomedical Science (Effective:1st July 2011) Kulliyyah of Nursing Kulliyyah of Nursing > Department of Basic Medical Sciences for Nursing
Depositing User:	Dr. Azmir Ahmad
Date Deposited:	18 Nov 2021 09:12
Last Modified:	03 Jun 2022 10:33
URI:	http://irep.iium.edu.my/id/eprint/93832

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