Total RNA was isolated from the harvested proximal stumps of sciatic nerve after injury and the collected NSCs using TRIzol reagent (Invitrogen, Carlsbad, CA). For the miRNA quantification, we used the TaqMan microRNA Reverse Transcription Kit and the TaqMan Universal Master Mix II with the TaqMan MicroRNA Assay of miRNAs (Applied Biosystems, Foster City, CA) to test the miR-7 expression level in the tissues or cells. The cdc42 levels were calculated relative to glyceraldehyde-3-phosphate dehydrogenase (internal control) via the 2−ΔΔCt method using a real-time polymerase chain reaction system according to the manufacturer’s instructions with SYBR® Green Master Mix (Applied Biosystems, Foster City, CA).
Taqman microrna assay of mirnas
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TaqMan MicroRNA Assay is a laboratory tool used for the detection and quantification of microRNA (miRNA) molecules. It provides a standardized and sensitive method for analyzing the expression levels of specific miRNAs in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (4 mentions)
Taqman universal master mix 2, by Thermo Fisher Scientific (4 mentions)
Sybr green master mix, by Thermo Fisher Scientific (3 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Rt reagent kit with gdna eraser, by Takara Bio (1 mentions)
Nanodrop nd2000, by Takara Bio (1 mentions)
Sybr green master mix, by Takara Bio (1 mentions)
4 protocols using taqman microrna assay of mirnas
1
Quantification of miR-7 and cdc42 in Nerve Cells
2
Quantification of miR-503 and bcl-2 Expression
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After undergoing various treatments, total RNA samples were extracted from the cultured cells using Trizol (Invitrogen) according to the manufacturer’s instructions. The Taq-Man microRNA Reverse Transcription Kit and the TaqMan Universal Master Mix II with the TaqMan MicroRNA Assay of miRNAs (Applied Biosystems, USA) were used to test the miR-503 expression levels of the miRNAs in the tissues and cells. The bcl-2 levels were calculated relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (internal control) via the 2−ΔΔCt method using a real-time PCR system according to the manufacturer’s instructions in SYBR® Green Master Mix (Applied Biosystems, USA).
3
Quantification of miRNA and mRNA Expression
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Total RNA samples were extracted from lung tissue or A549 cells using Trizol (Invitrogen) according to the manufacturer’s instructions. The Taqman microRNA Reverse Transcription Kit and Taqman Universal Master Mix II with the TaqMan MicroRNA Assay of miRNAs (Applied Biosystems, Foster City, USA) were used for testing the miR-144-3P expression level. The level of AQP1 and CASC2 was calculated relative to internal control using the 2−ΔΔCt method using real-time PCR system according to manufacturer’s instructions in SYBR green master mix (Applied Biosystems).
4
Quantitative Analysis of miRNA Expression
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Total RNA of cultured cells was isolated by TRIzol reagent (Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. RNA quantification was performed using a NanoDrop ND-2000 and cDNA was reversely transcribed using the RT reagent Kit with gDNA Eraser (Takara, Dalian, China) according to the manufacturer's protocols. Quantitative RT-PCR was performed using SYBR Green Master Mix (Takara, Dalian, China) on a Bio-Rad system. The expression level of miRNA was examined using Taqman microRNA Reverse Transcription Kit and Taqman Universal Master Mix II with the TaqMan MicroRNA Assay of miRNAs (Applied Biosystems, USA). GAPDH and U6 were used as the internal normalizer. The PCR reaction conditions for all of the assays were 94°C for 30 seconds, followed by 40 cycles of amplification (94°C for 5 seconds, 60°C for 30 seconds, and 72°C for 30 seconds). Relative mRNA level was calculated using the 2−ΔΔCt method. The sequences of primers were as follows: HOXA11-AS forward: 5'-CGGCTAACAAGGAGATTTGG-3' and reverse: 5'-AG- GCTCAGGGATGGTAGTCC-3'. miR-214-3p forward: 5'-ACAGCAGGCACAGACAGG-3' and reverse: 5'- GTGCAGGGTCCGAGGT-3'. GAPDH forward: 5'-GGGAGCCAAAAGGGTCAT-3' reverse: 5'- GAGTCCFTTCCACGATACCAA-3'.
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