Ribozol

Manufactured by Merck Group

Sourced in United States

Ribozol is a laboratory reagent used for the extraction and purification of RNA from biological samples. It functions as a guanidinium thiocyanate-phenol-chloroform solution, which effectively lyses cells and inactivates RNases to preserve the integrity of the extracted RNA.

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Lab products found in correlation

All in one mirna qrt pcr detection kit, by GeneCopoeia (2 mentions) Sybr green master mix, by Bio-Rad (1 mentions) Nanodrop 2000, by Thermo Fisher Scientific (1 mentions) Primescript rt reagent kit, by Takara Bio (1 mentions) Gdna eraser, by Takara Bio (1 mentions) All in one mirna rt qpcr detection kit, by GeneCopoeia (1 mentions) Sybr green rt pcr kit, by Vazyme (1 mentions) Qscript cdna supermix, by Quanta Biosciences (1 mentions) Qiaxcel advanced system, by Qiagen (1 mentions) Chloroform, by Merck Group (1 mentions) Ribozol, by Avantor (1 mentions) Qiaxcel rna qc kit v2, by Qiagen (1 mentions) Am9939, by Thermo Fisher Scientific (1 mentions) Purelink mirna isolation kit, by Thermo Fisher Scientific (1 mentions) Ssrt 4 system, by Thermo Fisher Scientific (1 mentions) Sybr green realtime pcr master mix, by Toyobo (1 mentions) Dna eraser, by Takara Bio (1 mentions) Dnase 1, by Thermo Fisher Scientific (1 mentions) Dnase 1, by Merck Group (1 mentions) Sybr green pcr kit, by Takara Bio (1 mentions)

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Ribozol reagent (3 citations)

11 protocols using ribozol

Quantifying Gene and miRNA Expression in SHP-77 Cells and Tissue Samples

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To measure gene expression levels, SHP-77 cells (10⁵ cells harvested at 24 h post-transfection) and tissue samples (0.03 g tissue ground in liquid nitrogen) were subjected to total RNA extractions using Ribozol (Sigma-Aldrich). To harvest miRNAs, 85% of ethanol was used to precipitate RNA samples. All RNA samples were treated with DNase I for 2 h at 37 °C to digest genomic DNAs.
To measure the levels of LUADT1 and Twist1 mRNA expression, the MMLV Reverse Transcriptase kit (Lucigen) was used to perform all reverse transcriptions and SYBR Green Master Mix (Bio-Rad) was used to prepare qPCR assays with GAPDH as an endogenous control.
To measure the levels of mature miR-15a-3p expression, poly (A) addition, reverse transcriptions and all qPCR assays were performed using All-in-One™ miRNA qRT-PCR Detection Kit (Genecopoeia). The endogenous control was U6.
Three replicate reactions were set for each experiment and Ct values were processed using the 2^-ΔΔCT method.

Wang D., Wu W., Huang W., Wang J., Luo L, & Tang D. (2019). LncRNA LUADT1 sponges miR-15a-3p to upregulate Twist1 in small cell lung cancer. BMC Pulmonary Medicine, 19, 246.

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Quantitative Analysis of miRNA and mRNA Levels

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Ribozol (Sigma-Aldrich; Merck KGaA) was used to isolate total RNAs from tissues and cells. miRNAs were harvested by precipitating RNAs with 85% ethanol. RNA concentrations were measured using NanoDrop 2000 (Thermo Fisher Scientific. Inc.) and genomic DNAs were digested using gDNA eraser (Takara Bio, Inc.). RNA samples with an OD 260/280 ratio ~2.0 (pure RNA) were reverse transcribed using the PrimeScript RT Reagent kit (Takara Bio, Inc.). The detailed workflow was as follows: 37°C For 15 min and 85°C for 5 sec then maintained at 4°C. Then the qPCR assays performed using SYBR-Green RT-PCR kit (Vazyme Biotech Co., Ltd.) to measure the expression levels of GABPB1-IT1 and PEDF mRNA. The detailed workflow was as follows: 95°C For 10 min, 40 cycles of 95°C for 15 sec and 60°C for 45 sec, final annealing at 72°C for 10 min. The expression levels of mature miR-93 were measured using the All-in-One™ miRNA RT-qPCR Detection kit (GeneCopoeia, Inc.). U6 and GAPDH were used as the endogenous controls of miR-93 and PEDF, respectively. PCR reactions were performed in triplicate and Cq values were processed using the Δ-Δq (ΔΔCq) method (18 (link)).

Li B., Wei Y., Ge Q., Duan Y, & Guo L. (2021). lncRNA GABPB1 intronic transcript 1 upregulates pigment epithelium-derived factor via miR-93 to suppress cell proliferation in hepatocellular carcinoma. Oncology Letters, 21(4), 260.

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RNA Extraction and cDNA Synthesis Protocol

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Tissue samples were ground in liquid nitrogen using mortar and pestle, followed by homogenization in Ribozol (N580-CA, Amresco) using a polytron homogenizer. Pelleted cultured cells were re-suspended in Ribozol. RNA was isolated by adding chloroform (C2432, Sigma) to the tissue/cell-Ribozol suspension following the manufacturer’s directions. The RNA was resuspended in 20 µL nuclease free water (AM9939, Ambion). RNA quality and quantity was assessed using a QIAxcel Advanced System (Qiagen) and QIAxcel RNA QC Kit v2.0 (Qiagen). From 1 µg of RNA as a template first-strand cDNA was synthesized using qScript cDNA supermix (CA101414-104, Quanta Biosciences). The reverse transcriptase reaction sequence consisted of incubation at 25 °C for 5 min, followed by incubation at 42 °C for 30 min and reverse transcriptase enzyme inactivation by incubation at 85 °C for 5 min. Resulting cDNA samples were stored at −20 °C until further analysis.

Perez L.J., Rios L., Trivedi P., D’Souza K., Cowie A., Nzirorera C., Webster D., Brunt K., Legare J.F., Hassan A., Kienesberger P.C, & Pulinilkunnil T. (2017). Validation of optimal reference genes for quantitative real time PCR in muscle and adipose tissue for obesity and diabetes research. Scientific Reports, 7, 3612.

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Extraction and Purification of Total RNAs

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Isolation of total RNAs from OC tissues, non-tumor tissues, and UWB1.289 cells was performed using Ribozol (Sigma-Aldrich). RNA precipitation was performed using 85% ethanol to harvest miRNAs. Genomic DNA was removed by DNase I digestion at 37 °C for 2 h.

Cui K, & Zhu G. (2020). LncRNA CTBP1-AS2 regulates miR-216a/ PTEN to suppress ovarian cancer cell proliferation. Journal of Ovarian Research, 13, 84.

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Total RNA and miRNA Extraction

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Ribozol (Sigma-Aldrich) was used to extract total RNAs from CSCC and non-tumor tissues as well as CSCC cells. PureLink miRNA Isolation Kit (Thermo Fisher Scientific) was used to extract miRNAs. DNA-Eraser Genomic DNA Removal Kit (FroggaBio) was used to digest all RNA samples to remove genomic DNA.

Lan L., Liang Z., Zhao Y, & Mo Y. (2020). LncRNA MCM3AP-AS1 inhibits cell proliferation in cervical squamous cell carcinoma by down-regulating miRNA-93. Bioscience Reports, 40(2), BSR20193794.

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Quantification of miRNA and mRNA Expression

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Ribozol (Sigma-Aldrich) was used to perform all RNA extractions following the manufacture’s instructions. RNA samples were precipitated using 85% ethanol to harvest miRNAs. All RNA samples were digested by DNA eraser (Takara, USA). SSRT IV system (Thermo Fisher Scientific) was used to transcribe total RNAs into cDNAs. With cDNA samples as template, SYBR® Green Realtime PCR Master Mix (Toyobo, Japan) was used to prepare all qPCR mixtures. GAPDH was used as the endogenous control to measure the expession levels of MIR17HG and Bach-1. All-in-One™ miRNA qRT-PCR Detection Kit (GeneCopoeia) was used to measure the expression levels of mature miR-142-3p. All steps including polyadenylation, reverse transcriptions and qPCR assays were performed following the instructions from GeneCopoeia. U6 was used as the endogenous control. All qPCR assays were repeated 3 times and data normalizations were performed using 2^-ΔΔCq method.

Wei S., Liu J., Li X, & Liu X. (2020). LncRNA MIR17HG inhibits non-small cell lung cancer by upregulating miR-142-3p to downregulate Bach-1. BMC Pulmonary Medicine, 20, 78.

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Comprehensive RNA Extraction Protocol

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H1993 cells were harvested and counted. All tissue samples (about 0.012 g) were ground into powder in liquid nitrogen. Total RNAs in 4 × 10⁵ cells or tissue samples were extracted using Ribozol (Sigma-Aldrich, USA). To harvest all types of RNAs (such as miRNAs), RNAs were precipitated and washed using 85% ethanol.

Huang S., Li C., Huang J., Luo P., Mo D, & Wang H. (2020). LncRNA FEZF1-AS1 promotes non-small lung cancer cell migration and invasion through the up-regulation of NOTCH1 by serving as a sponge of miR-34a. BMC Pulmonary Medicine, 20, 110.

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miRNA Extraction from Cell and Tissue

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H23 cells were harvested at 48h post-transfection and cells were counted. Tissue samples (0.04g per sample) were ground in liquid nitrogen to make a fine powder. Total RNAs in cells and tissues were extracted using Ribozol (Sigma-Aldrich). Eighty-five percent ethanol was used to precipitate and wash RNA samples to harvest miRNAs.

Li C., Lei Z., Peng B., Zhu J, & Chen L. (2020). LncRNA HCP5 Stimulates the Proliferation of Non-Small Cell Lung Cancer Cells by Up-Regulating Survivin Through the Down-Regulation of miR-320. Cancer Management and Research, 12, 1129-1134.

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Extraction and Purification of Total RNA

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Total RNAs were extracted from U2OS and MG-63 cells (10⁸ cells) and paired tissue samples (0.02 g) using Ribozol (Sigma-Aldrich). To remove genomic DNAs, all RNA samples were digested with DNase I (Invitrogen) at 37 °C for 2 h. Urea-PAGE gel (5%) was used to evaluate RNA integrity.

He P., Xu Y, & Wang Z. (2020). LncRNA SNHG10 increases the methylation of miR-218 gene to promote glucose uptake and cell proliferation in osteosarcoma. Journal of Orthopaedic Surgery and Research, 15, 353.

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Hypoxia-Induced Gene and miRNA Expression

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RNAs isolation was performed on plasma and HRPTEpCs using Ribozol (Sigma-Aldrich). For hypoxia treatment, HRPTEpCs were cultured under hypoxia conditions (5% CO 2 /94% N 2 /1% O 2 ) for 12, 24 and 48 h prior to use. Subsequently, RNA was precipitated using 85% ethanol. Genomic DNA was eliminated by treating RNA samples with DNase I (Sigma-Aldrich) at 37Â°C for 2 h. RNA concentrations were measured using a NanoDrop 2000 Spectrophotometer (Thermo Scientific). qPCR assays were conducted using the Precision nanoScript2 Reverse Transcription Kit (PrimerDesign) and SYBR Green PCR Kit (Takara Bio). GAPDH was used as the endogenous control. All-in-Oneâ"¢ miRNA qRT-PCR Detection Kit (Genecopoeia) was used to detect the expression of mature miR-204-5p. Gene expression was normalized using the 2 âˆ'Î"Î"Ct method. PCR reactions were replicated for 3 times.

Feng F., Zhang R, & Long L. (2024). LncRNA GABPB1-IT1 Is Upregulated in Ischemia-Induced Acute Kidney Injury and Downregulates miR-204-5p to Promote Hypoxia-Induced Human Renal Proximal Tubular Epithelial Cell Apoptosis. Kidney & blood pressure research, 49(1).

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