We then confirmed miRNA expression of selected miRNA that controlled multiple genes and/or controlled pathways which are known to be important in vascular calcification, by real time PCR as previously described [17 (link)] using TaqMan miRNA assays (Applied Biosystems, Foster City, CA). Forty ng of total RNA were used for reverse transcription to synthesize complementary DNA using TaqMan miRNA-specific primers and the Taq reverse transcription kit (Applied Biosystems, Foster City, CA). Target-specific PCR primers (miR-667, miR-702, miR-3562, miR-3568 and miR-3584) were obtained from Applied Biosystems. Real-time PCR amplifications were performed using TaqMan miRNA assays (TaqMan MGP probes, FAM dye-labeled) using Applied Biosystems 7500 Real-Time PCR systems (Applied Biosystems). The cycle number at which the amplification plot crosses the threshold was calculated (CT), and the ΔΔCT method was used to analyze the relative changes in gene expression and normalized by U6, a non-human ubiquitous miRNA.
Target specific pcr primers
Manufactured by Thermo Fisher Scientific
Sourced in United States
Target-specific PCR primers are short DNA sequences designed to selectively amplify a specific region of a target DNA molecule during the Polymerase Chain Reaction (PCR) process. These primers serve as the starting points for DNA synthesis, allowing for the selective and exponential amplification of the targeted genetic sequence.
Automatically generated - may contain errors
2 protocols using target specific pcr primers
1
Validation of miRNA Expression in Vascular Calcification
2
Aortic Gene Expression Analysis
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