Trizol rna extraction reagent

Manufactured by Takara Bio

Sourced in China

Trizol is a reagent used for the extraction and purification of RNA from biological samples. It is a single-phase solution composed of phenol and guanidine isothiocyanate, which facilitates the lysis of cells and the separation of RNA from DNA and proteins. The reagent allows for the efficient and reliable isolation of high-quality RNA for various downstream applications, such as gene expression analysis, Northern blotting, and RT-PCR.

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Lab products found in correlation

Cell cycle assay kit, by Beyotime (1 mentions) Sybr green pcr master mix, by Takara Bio (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Reverse transcription kit, by Toyobo (1 mentions) Sybr premix ex taq kit, by Takara Bio (1 mentions) One step primescript rt pcr kit, by Takara Bio (1 mentions) Cfx96 real time pcr detection system, by Bio-Rad (1 mentions) Abi 7300 real time pcr system, by Takara Bio (1 mentions) Primescript rt master mix, by Takara Bio (1 mentions)

4 protocols using trizol rna extraction reagent

Lentiviral Knockdown of ZYX and OGT

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For RNA interference, lentiviral packaging plasmids psPAX2 and pMD2.G were cotransfected with the PLKO.1 backbone plasmid into HEK293T cells to produced virus. The following targeting sequences were inserted into the pLKO.1 vector for indicated gene: ZYX, 5’-AGAAGGTGAGCAGTA TTGATT-3’; OGT, 5’-TGCACAATCCTGATAAATTTGA-3’. We used nontarget sequence as negative shRNA control. The sequence is 5’-CAACAAGATGAAGAGCACCAA-3’.
For RT–qPCR, total RNA was extracted using the Trizol RNA extraction reagent (TaKaRa; catalog no.: 9109). Complementary DNA was reverse transcripted using the HIScriptII One Step RT–qPCR Kit (Vazyme; catalog no.: R223-01). Gene expression was analyzed by real-time qPCR using the SYBR Green quantitative PCR Mix (Vazyme; catalog no.: Q311-00). The relative mRNA abundance was calculated by normalization to ACTB mRNA. The following primers were used for RT–qPCR: PUMA (5’-GACCTCAACGCACAGTACGAG-3′ and 5’-AGGAGTCCCATGATGAGATTGT-3′); ACTB (5’-CTCCTTAATGTCACGCACGAT-3′ and 5’-CATGTACGTT GCTATCCAGGC-3′).

Zhao Y., Yue S., Zhou X., Guo J., Ma S, & Chen Q. (2022). O-GlcNAc transferase promotes the nuclear localization of the focal adhesion–associated protein Zyxin to regulate UV-induced cell death. The Journal of Biological Chemistry, 298(4), 101776.

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Regulation of Cyclin D1 in Thyroid Cancer

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Human PTC cells (TPC-1) (BNCC, Beijing, China); cattle TSH and TH (Sigma, San Francisco, CA, USA); methyl thiazolyl tetrazolium (MTT) (Sigma); RPMI-1640 medium and fetal calf serum (Gibco, Grand Island, NY, USA); cell cycle assay kit (Beyotime, Shanghai, China); TRIzol RNA extraction reagent (Takara, Shiga, Japan); reverse transcription kit (Toyobo, Osaka, Japan); human cyclin D1 and β-actin primers (Shanghai Sangon Biomedical Engineering Co., Ltd., Shanghai, China); SYBR Green PCR Master mix (Takara); cyclin D1 enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Inc., Minneapolis, MN, USA).

Ma Y., Zhang X, & Wang Y. (2017). Reactivity of thyroid papillary carcinoma cells to thyroid stimulating hormone-dominated endocrine therapy. Oncology Letters, 14(6), 7405-7409.

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Quantitative Analysis of MEG3 Expression

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The total RNA was extracted from PT-U cells and embryonic villi using the Trizol RNA extraction reagent (TaKaRa, Otsu, Japan), and reverse transcribed using the Prime-Script™ one-step RT-PCR kit (TaKaRa). The resulting cDNAs were used as a template for real-time quantitative PCR (qPCR) assay using SYBR Premix Ex Taq kit (TaKaRa), as per instructions provided in the kit, on the CFX96 Real-Time PCR Detection System (Bio-Rad, Hercules, California, USA). The relative expression level of MEG3 was analyzed using the 2^−ΔΔCt method and normalized using GAPDH as the internal reference gene. The primer sequences are listed in Additional file 2: Table S2.

Zhang J., Liu X, & Gao Y. (2021). The long noncoding RNA MEG3 regulates Ras-MAPK pathway through RASA1 in trophoblast and is associated with unexplained recurrent spontaneous abortion. Molecular Medicine, 27, 70.

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Osteogenic Differentiation Gene Expression

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After 7 days' culture, the total RNA was extracted using Trizol RNA extraction reagent (TAKARA, Dalian, Liaoning, China). cDNA was synthesized with PrimeScript RT Master Mix (TAKARA). RT-PCR was performed by ABI 7300 Real-Time PCR System using Premix Ex Tag (TAKARA). The cycling parameters used were as follows: denaturation at 95°C for 15 min and 30 amplification cycles (15 s at 95°C, 25 s at 60°C, and 10 s at 72°C). The primers were RUNX2, COL1A2, osteopontin (OPN), and osteocalcin (OCN) (Table 1). Expression of β-actin was used as an internal control.

Yin L., Cheng W., Qin Z., Yu H., Yu Z., Zhong M., Sun K, & Zhang W. (2015). Effects of Naringin on Proliferation and Osteogenic Differentiation of Human Periodontal Ligament Stem Cells In Vitro and In Vivo. Stem Cells International, 2015, 758706.

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