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- W2468769003 abstract "Blood samples are collected and stored to explore feasible biomarkers in most dementia care registry study. In recent years, examining potential plasma miRNAs are considered less invasive and might provide an opportunity to identify candidate biomarker for Alzheimer's disease (AD). However, it remains a challenge to detect miRNAs from longer preserved blood samples. We aimed to explore an approach to extract, separate and detect the plasma miRNAs from the preserved blood samples. Preserved plasma and blood cell samples were separated by ultracentrifugation. Total RNA was isolated from these samples using the TRIzol Reagent. Then, miRNAs in the total RNA were reversely translated into cDNA, and the relative quantitative expression of these miRNAs was detected by qPCR. Prior to the validation of miRNA qRT-PCR of human blood samples, the procedures of routine PCR, cDNA aPCR, miRNA-PCR of mouse brain tissues, and miRNA qRT-PCR of mouse blood samples were sequentially validated. Then, a standard procedure of human blood miRNA qRT-PCR was developed and optimized. As shown in Figure 1a, the miRNA in preserved blood samples (plasma, whole blood and blood cells) have be successfully amplified as compared with the negative control sample (H2O) and the positive control sample (mouse brain tissues). There was no significant difference in the time to reach the Threshold fluorescence among miRNAs from preserved whole blood, plasma and blood cells (Figure 1a). The Ct values of miRNA in plasma, whole blood and blood cells were in the same level (Figure 1b). With the total RNA extracted and separated with this protocol, it was found that miR-770-3p was significantly up-regulated in the 5XFAD mouse model while miR-1264-3p, miR-1298-5p, and miR-448-3p were down-regulated (Figure 2)." @default.
- W2468769003 created "2016-07-22" @default.
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- W2468769003 date "2015-07-01" @default.
- W2468769003 modified "2023-10-16" @default.
- W2468769003 title "P1-008: A protocol for quantifying the expression of mirnas from preserved human blood" @default.
- W2468769003 doi "https://doi.org/10.1016/j.jalz.2015.06.204" @default.
- W2468769003 hasPublicationYear "2015" @default.
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