For microRNA quantification of miR-329-3p and miR-495-3p, RNA was reversed transcribed using the TaqmanTM MicroRNA Reverse-Transcription Kit (Thermo Fisher Scientific) and, subsequently, quantified using microRNA-specific TaqmanTM qPCR kits (Thermo Fisher Scientific) on the VIIa7 (Thermo Fisher Scientific). MicroRNA expression was normalized against U6 small nuclear RNA.
Taqman qpcr kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
TaqmanTM qPCR kits are a set of reagents used for quantitative real-time polymerase chain reaction (qPCR) analysis. The kits include primers, probes, and enzymes necessary to perform qPCR experiments.
Automatically generated - may contain errors
4 protocols using taqman qpcr kit
1
Quantifying miR-329-3p and miR-495-3p
2
MicroRNA Quantification of Specific Targets
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For microRNA quantification of miR-433-3p, miR-494-3p, and miR-495-3p, in all samples, RNA was reversed transcribed using the TaqmanTM MicroRNA Reverse Transcription Kit (Thermo Fisher Scientific) and subsequently quantified using microRNA-specific TaqmanTM qPCR kits (Thermo Fisher Scientific) on the VIIa7 (Thermo Fisher Scientific). MicroRNA expression was normalized against U6 small nuclear RNA.
3
Quantitative RNA Expression Analysis in Rat Substantia Nigra
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TRIzol extraction of total RNA from rat substantia nigra tissue (100 mg) was performed according to the instructions of the kit. A total of 20 µl reaction system was prepared and cDNA was synthesized according to the following conditions: 37°C for 15 min and 85°C for 5 sec. A total of 2 µl of reverse transcription product was used for the PCR reaction according to the instructions of TaqMan qPCR kit (Thermo fisher, Waltham, MA, USA) and GAPDH was used as endogenous control. Reaction conditions were: 95°C for 60 sec, followed by 40 cycles of 95°C for 10 sec and 60°C for 30 sec. This experiment was performed in triplicate manner and data were processed using 2−ΔCq method (14 (link)). Primer sequences are listed in Table I .
4
Quantitative Analysis of Inflammatory and Remodeling Markers in Murine Aorta
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RNA was isolated from aortas and SMCs using Trizol-reagent (Invitrogen) and RNeasy columns (QIAGEN). RNA (1 μg) was used for reverse transcription with SuperScript III reverse transcriptase (Invitrogen). The reaction was topped up to 200 μl with water, and 2 μl were used for quantitative real-time PCR reaction with TaqMan qPCR Kit (Thermo Fischer Scientific) from Eurogentec. Standardization was performed with primers Tata-box protein. VCAM-1; Mm00449197_m1, ICAM-1; Mm00516023_m1, MCP-1; Mm00441242_m1, MCP-3; Mm00443113_m1, MIF; Mm01611157_gH, Fractalkine; Mm00436454_m1, KC; Mm00433859_m1, Eotaxin; Mm00441238_m1, IL-6; Mm00446190_m1, IL-1β; Mm00434228_m1, TGFβ; Mm03024053_m1, IL-10; Mm00439616_m1, TNF; Mm00443258_m1, Fas; Mm00433237_m1, MMP-2; Mm00439498_m1, MMP-3; Mm00440295_m1, MMP-9; Mm00442991_m1, MMP-13; Mm00439491_m1, Tata-box protein; Mm01277042_m1.
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