Triagent

Manufactured by Molecular Research Center

Sourced in United States

Triagent is a multi-purpose laboratory instrument designed for nucleic acid extraction and purification. It utilizes a unique centrifugation-based method to efficiently separate and purify DNA, RNA, and proteins from a variety of sample types.

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Lab products found in correlation

Genespring gx software 14 9, by Agilent Technologies (1 mentions) Feature extraction software 10, by Agilent Technologies (1 mentions) Bioanalyzer 2100 system, by Agilent Technologies (1 mentions) Superscript 4 first strand synthesis system, by Thermo Fisher Scientific (1 mentions) Power sybr green master mix, by Thermo Fisher Scientific (1 mentions) Quantstudio 7 flex real time pcr system, by Thermo Fisher Scientific (1 mentions) Real time detector system, by Thermo Fisher Scientific (1 mentions) Sybr green pcr mixture, by Takara Bio (1 mentions)

3 protocols using triagent

Microarray Analysis of Neuro2a Cell miRNA

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For the microarray analysis, total RNA (miRNA) was isolated from the Neuro2a cells using Triagent (Molecular Research Center, Cincinnati, OH, USA), and the quality was evaluated by a 2100 Bioanalyzer system (Agilent Technologies, Santa Clara, CA, USA). Cy3-labeled probes were prepared from 200 ng of total RNA using the Low Input Quick Amp Labeling Kit, one-color, (Agilent Technologies, #5190-2305) and hybridized with a microarray slide (Whole Mouse Genome DNA microarray 4x44K Ver. 2.0; Agilent Technologies) for 17 h at 65 °C. The microarray slide was washed and scanned with a microarray scanner (ArrayScan, Agilent Technologies) to obtain the fluorescent signal of the probes. The signal was processed for digitization using Feature Extraction software 10.7 (Agilent Technologies) and analyzed with GeneSpring GX software 14. 9 (Agilent Technologies) for gene expression.

Oh-hashi K., Nakamura H., Ogawa H., Hirata Y, & Sakurai K. (2023). Elucidation of OSW-1-Induced Stress Responses in Neuro2a Cells. International Journal of Molecular Sciences, 24(6), 5787.

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Validating miR-187b and Target Gene Expression

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Quantitative PCR (qPCR) was used to validate predicted differential expression for the miR-187b and its putative “hub” target genes in murine hearts. Total RNA from whole heart tissue was isolated using Tri-Agent (Molecular Research Center Inc) and reverse transcribed to cDNA using the “SuperScript IV First-Strand Synthesis System” (Thermofisher). For each hub gene following primers were designed. Sequence can be found in the Supplemental table 1. The primers for miR-187-5p and miR-187-3p were designed as previously described (Busk, 2014) (23 ). qPCR was performed with Power SYBR Green Master Mix (Applied Biosystems) using the QuantStudio 7 Flex Real-Time PCR System (Applied Biosystems). The average threshold count (Ct) value of three technical replicates was used in all calculations. β-actin and U6 were used as loading controls for mRNA and miRNA targets respectively because they displayed the lowest standard deviation among groups compared to other housekeeping genes tested. Data were analyzed using the 2−ΔCt method depicted by Schmittgen and Livak 2008 (24 (link)). Relative mRNA data are expressed as mean ± standard deviation.

Ektesabi A.M., Mori K., Tsoporis J.N., Vaswani C.M., Gupta S., Walsh C., Varkouhi A.K., Mei S.H., Stewart D.J., Liles W.C., Marshall J.C., Hu P., Parker T.G, & dos Santos C.C. (2020). Mesenchymal Stem/Stromal Cells Increase Cardiac miR-187-3p Expression in a Polymicrobial Animal Model of Sepsis. Shock (Augusta, Ga.), 56(1), 133-141.

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Quantitative RT-PCR from HeLa Cells

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Total RNA from HeLa cells was prepared using the TRI agent (Molecular Research Center, Cincinnati, OH, USA) according to the manufacturer's instructions and then reverse transcribed to generate complementary DNA. Quantitative RT-PCR was performed using the SYBR Green PCR mixture (Takara Bio Inc., Shiga, Japan) and a real-time detector system (Applied BioSystems, Foster City, CA USA). GAPDH level was used to normalize transcript levels.

Kim Y.S., Kim S.H., Shin J., Harikishore A., Lim J.K., Jung Y., Lyu H.N., Baek N.I., Choi K.Y., Yoon H.S, & Kim K.T. (2014). Luteolin Suppresses Cancer Cell Proliferation by Targeting Vaccinia-Related Kinase 1. PLoS ONE, 9(10), e109655.

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