Rsc simply rna tissue kit

Manufactured by Promega

The Promega RSC) simply RNA Tissue kit is a laboratory product designed for the isolation of high-quality RNA from a variety of tissue samples. It provides a simple and efficient method for extracting RNA without the use of organic solvents.

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Lab products found in correlation

Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Cf96 real time system, by Bio-Rad (1 mentions) Maxwell rsc instrument, by Promega (1 mentions) Ncounter pancancer immune profiling panel, by NanoString (1 mentions) Agilent tapestation rna screentape, by Agilent Technologies (1 mentions) Quant it broad range rna assay kit, by Thermo Fisher Scientific (1 mentions) Applied biosystems high capacity cdna archive kit, by Thermo Fisher Scientific (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Rq1 rnase free dnase kit, by Promega (1 mentions)

4 protocols using rsc simply rna tissue kit

Aortic Expression Analysis of KDM5C and KDM6A

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RNA was extracted from the thoracic aorta (defined as the segment extending from the aortic root to the diaphragm) and abdominal aorta (defined as the segment extending from the diaphragm to the iliac bifurcation) from XXF and XOF Ldlr^−/− mice fed a Western diet for 1 week. RNA was extracted using a Maxwell Rapid Sample Concentrator (RSC) simply RNA Tissue kit (REF#AS1340, Promega, Madison, WI). RNA (226 ng) was reverse transcribed to cDNA using Supermix (cat#95048-500, Quanta Biosciences, Gaithersburg, MD). cDNA was diluted (1:10) and mRNA abundance of Kdm5c or Kdm6a was quantified by RT-PCR using SYBER Green FastMix (cat#95071-012, Gaithersburg, MD) on a BioRad quantitative real-time PCR thermocycler (CF96 Real-Time system, BioRad, Hercules, CA). mRNA abundance was determined using the ΔΔCt method and normalized to β-actin as a housekeeping gene.
Primer sequences for Kdm5c were: Forward 5’-ACCCACCTGGCAAAAACATTGG-3’; reverse 5’-ACTGTCGAAGGGGGATGCTGTG-3’. Primers sequences for β-actin were: 5’-GCTCTGGCTCCTAGCACCAT-3’; reverse 5’-GCCACCGATCCACACAGAGT-3’. Primer sequences for Kdm6a were: Forward 5’-CCAATCCCCGCAGAGCTTACCT-3’; reverse 5’-TTGCTCGGAGCTGTTCCAAGTG-3’.

AlSiraj Y., Thatcher S.E., Blalock E., Saintilnord W.N., Daugherty A., Lu H.S., Luo W., Shen Y.H., LeMaire S.A., Arnold A.P, & Cassis L.A. (2020). Monosomy X in Female Mice Influences the Regional Formation and Augments the Severity of Angiotensin II-induced Aortopathies. Arteriosclerosis, thrombosis, and vascular biology, 41(1), 269-283.

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Immune and Metabolic Gene Expression Analysis

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Whole-blood stored in PAXgene Blood RNA (BD Biosciences) tubes was thawed and RNA was isolated using the automated Maxwell RSC Instrument (Promega) and the RSC simplyRNA tissue kit (Promega). Isolated RNA was analysed using both the commercially available nCounter PanCancer Immune Profiling Panel consisting of 770 genes (Nanostring Technologies) and a custom panel targeting 48 metabolic and immune genes (Nanostring Technologies). A complete list of the genes assessed in the custom panel can be found in Supplementary Table 1. Assays were completed according to manufacturers’ instructions. Samples were processed using the automated nCounter Prep Station prior to direct digital counting on the nCounter Digital Analyser platform (Nanostring Technologies) at a field of view of 555. Transcript counts were normalized according to the geomean of the housekeepers for each panel. Genes with average counts lower than the background (approximately 20) were removed. Genes that exhibited significant differences between groups on Student’s t-test (P<0.05) or that exhibited a 1.5-fold change difference were retained for downstream between-group analysis.

Hatton-Jones K.M., West N.P., Thang M.W., Chen P.Y., Davoren P., Cripps A.W, & Cox A.J. (2024). Gut Microbiome and Metabolic and Immune Indices in Males with or without Evidence of Metabolic Dysregulation. Journal of Obesity & Metabolic Syndrome, 33(1), 64-75.

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Automated RNA Isolation and NanoString Analysis

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RNA was isolated from spinal cord tissue using automated Promega Maxwells with the RSC simplyRNA Tissue Kit as per the manufacturer’s recommendations (Promega, cat #AS1340). RNA quality was assessed using Agilent Tape Station RNA ScreenTape (Agilent, cat #5067-5576) and quantitated by Invitrogen Quant-iT RNA Broad Range Assay Kit (Invitrogen, cat #Q10213). 150 ng of RNA was used as input for NanoString Autoimmune Panel (NanoString cat #XT-CSO-MAIP1-12) according to nCounter XT CodeSet Gene Expression Assays protocol. Resulting RCC files were examined with NanoString nSolver software for QC analysis. Briefly, samples had a background threshold set to 25 counts and normalized to a set of housekeeping genes with low variance (%CV < 40%) and medium or high counts (>50).

de Picciotto S., DeVita N., Hsiao C.J., Honan C., Tse S.W., Nguyen M., Ferrari J.D., Zheng W., Wipke B.T, & Huang E. (2022). Selective activation and expansion of regulatory T cells using lipid encapsulated mRNA encoding a long-acting IL-2 mutein. Nature Communications, 13, 3866.

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Hepatic Tissue RNA Extraction and Analysis

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Total RNA was extracted from ~15 mg of hepatic tissue using the RSC simply RNA Tissue Kit (Promega Corp., Madison, WI) and then purified using the RQ1 RNase-Free DNase Kit (Promega Corp.). Purity, concentration, and integrity of mRNA were evaluated using a NanoDrop 1000 spectrophotometer (Thermo Scientific, Wilmington, DE) and an Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA) system. All samples had a RNA integrity number >6. Conversion to cDNA was performed using the Applied Biosystems High Capacity cDNA Archive Kit (Applied Biosystems, Foster City, CA).

Gualdrón-Duarte L.B, & Allen M.S. (2018). Fuels derived from starch digestion have different effects on energy intake and metabolic responses of cows in the postpartum period. Journal of dairy science, 101(6).

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