Tri reagent extraction kit

Manufactured by Merck Group

Sourced in United States

The Tri Reagent extraction kit is a laboratory product developed by Merck Group. The kit is designed to facilitate the extraction and isolation of RNA, DNA, and proteins from various biological samples. It utilizes a single-step liquid-phase separation technique to effectively separate the biomolecules. The kit provides a streamlined and efficient method for researchers and scientists to obtain high-quality nucleic acids and proteins for further analysis and experimentation.

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Lab products found in correlation

Rq1 rnase free dnase, by Promega (2 mentions) Cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) μdrop plate, by Thermo Fisher Scientific (1 mentions)

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Tri-Reagent extraction TRI reagent kit TRI Reagent TRI kit

4 protocols using tri reagent extraction kit

Purification and Characterization of phiYY Virions

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phiYY virions were purified as previously described. Briefly, phiYY was inoculated into the log phase host in LB medium and cultured with aeration at 37 °C for 6 h. The phage lysate was centrifuged for 10 min at 10,000 × g and passed through a 0.22-μm filter. The phages were further purified by PEG8000 precipitation.
The phage dsRNA genome was isolated from the purified phage particles with the Tri Reagent extraction kit (Sigma-Aldrich). RNA integrity and size distribution were assessed on a 1% (w/v) agarose gel and visualized with ethidium bromide.

Yang Y., Lu S., Shen W., Zhao X., Shen M., Tan Y., Li G., Li M., Wang J., Hu F, & Le S. (2016). Characterization of the first double-stranded RNA bacteriophage infecting Pseudomonas aeruginosa. Scientific Reports, 6, 38795.

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RNA Extraction and cDNA Synthesis

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Using the Tri-Reagent™ extraction kit (Sigma-Aldrich, St. Louis, MO, USA) and following the given instructions, the total RNA from RAW 264.7 cells was extracted. The purity of the extracted RNA was evaluated using the μDrop Plate (Thermo Scientific). Next, RNA samples were diluted (1 μg μL^− 1). For the purpose of synthesizing first-strand cDNA, another kit was used (prime Script™) (TaKaRa BIO INC, Japan) following the manufacturer instructions. Synthesized cDNA was stored at − 80 °C.

Jayawardena T.U., Asanka Sanjeewa K.K., Shanura Fernando I.P., Ryu B.M., Kang M.C., Jee Y., Lee W.W, & Jeon Y.J. (2018). Sargassum horneri (Turner) C. Agardh ethanol extract inhibits the fine dust inflammation response via activating Nrf2/HO-1 signaling in RAW 264.7 cells. BMC Complementary and Alternative Medicine, 18, 249.

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Gene Expression Analysis in Larval Tissues

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Total RNA was isolated from 10 wandering third-instar larvae through use of a Tri Reagent extraction kit (Sigma-Aldrich, USA). RNA samples were treated with RQ1 RNase-Free DNase (Promega Corporation, USA). Reverse transcription was carried out with a Fermentas cDNA synthesis kit. Samples of cDNA were normalized to the rpL17A level, and the pro-apoptotic and proteasome subunit gene expressions were then determined by 20–25-cycle PCRs with the use of exon specific primers.

Kovács L., Nagy O., Pál M., Udvardy A., Popescu O, & Deák P. (2015). Role of the Deubiquitylating Enzyme DmUsp5 in Coupling Ubiquitin Equilibrium to Development and Apoptosis in Drosophila melanogaster. PLoS ONE, 10(3), e0120875.

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Quantitative RT-PCR Analysis of Gene Expression

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Total RNA was extracted from 10 larvae using the Tri Reagent extraction kit (Sigma-Aldrich, USA). RNA samples were treated with RQ1 RNase-free DNase (Promega Corporation, USA). The Thermo Fisher Scientific cDNA synthesis kit was used for reverse transcription, according to the manufacturer’s instruction. Samples were normalized to the rpL17A level using the following primers: RpL17A forward 5′-GAGCCAAGAACCTGTACG-3′ and reverse 5′-CAGGCATGACCTTCTTCC-3′. Gene expressions were determined using 20-25-cycle PCRs with exon-specific primers. The RT-PCR was quantified by using Fiji Image J (Supplementary Fig. 2).

Páhi Z.G., Kovács L., Szűcs D., Borsos B.N., Deák P, & Pankotai T. (2022). Usp5, Usp34, and Otu1 deubiquitylases mediate DNA repair in Drosophila melanogaster. Scientific Reports, 12, 5870.

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