Quantifluor rna system

Manufactured by Promega

Sourced in United States, Japan

The QuantiFluor RNA System is a fluorescence-based nucleic acid quantification tool. It allows for the accurate measurement of RNA concentrations in samples. The system utilizes a fluorescent dye that selectively binds to RNA, enabling precise quantification.

Automatically generated - may contain errors

Lab products found in correlation

Quantus fluorometer, by Promega (12 mentions) Quantifluor one dsdna system, by Promega (7 mentions) Rneasy mini kit, by Qiagen (5 mentions) Fragment analyzer, by Agilent Technologies (5 mentions) Maxwell rsc instrument, by Promega (4 mentions) Trizol, by Thermo Fisher Scientific (4 mentions) Turbo dna free kit, by Thermo Fisher Scientific (4 mentions) Nanodrop 2000, by Thermo Fisher Scientific (4 mentions) Magna pure compact, by Roche (4 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (4 mentions) Qubit fluorometer, by Thermo Fisher Scientific (4 mentions) Nanodrop spectrophotometer, by Thermo Fisher Scientific (4 mentions) 2100 bioanalyzer, by Agilent Technologies (3 mentions) Rna 6000 nano kit, by Agilent Technologies (3 mentions) Rneasy plus mini kit, by Qiagen (3 mentions) Bioanalyzer, by Agilent Technologies (3 mentions) Maxwell rsc simplyrna tissue kit, by Promega (3 mentions) Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (3 mentions) Trizol reagent, by Thermo Fisher Scientific (3 mentions) Trizol ls, by Thermo Fisher Scientific (2 mentions)

88 protocols using quantifluor rna system

Liver Total RNA Extraction and RT-qPCR

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA extraction was performed from liver samples using a total RNA isolation kit (JenaBioscience, Jena, Germany), following the manufacturer’s instructions. RNA quantification and purity was revealed using the QuantiFluor RNA System (Promega, Madison, WI, USA) and Quantus Fluorometer (Promega, Madison, WI, USA).
Furthermore, for cDNA synthesis from RNA, first-strand cDNA was synthesized with 1 ug of total RNA using a Revert Aid First-strand cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer′s instructions. These samples were then frozen at −80 °C until real-time PCR was used to determine gene expression.

Alzu’bi A., Zoubi M.S., Al-Trad B., AbuAlArjah M.I., Shehab M., Alzoubi H., Albals D., Abdelhady G.T, & El-Huneidi W. (2022). Acute Hepatic Injury Associated with Acute Administration of Synthetic Cannabinoid XLR-11 in Mouse Animal Model. Toxics, 10(11), 668.

+ Open protocol

+ Expand

Formulation and Characterization of mRNA-LNP

Check if the same lab product or an alternative is used in the 5 most similar protocols

mRNAs were diluted in 50 mM citrate buffer (pH 3.0) as the aqueous phase. Lipids were prepared by dissolving MC3-lipid, DSPC, cholesterol, PEG2000-DMG in ethanol at molar ratios of 50:10:38.5:1.5 (total concentration 12.5 mM). The lipids mixture was then mixed with the aqueous phase (mRNA) using the NanoAssemblr Benchtop (Precision #NIT0046) in compatible microfluidic cartridges. The formulated mRNA-LNP solution was first dialyzed against 1X PBS (pH ∼7) at 4°C overnight, and then concentrated by ultrafiltration using the Amicon Ultra Centrifugal Filter Unit (Millipore). The encapsulation efficiency and concentration of mRNAs were determined by the QuantiFluor RNA system (Promega #E3310). The size of mRNA-LNP particles was determined by dynamic light scattering (DLS) using samples diluted in PBS. DLS was performed using the Malvern Zetasizer (Malvern).

Zhang M., Singh N., Ehmann M.E., Zheng L, & Zhao H. (2023). Incorporation of noncanonical base Z yields modified mRNA with minimal immunogenicity and improved translational capacity in mammalian cells. iScience, 26(10), 107739.

+ Open protocol

+ Expand

RNA Extraction and Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Laboratories could use any RNA extraction method of their choice. The organising laboratory recommended a manual organic method [1] and a silica column based method (RNeasy® Mini Kit, Qiagen, Venlo, NL). For laboratories using a PGM/S5, it was highly recommended to use the organic extraction method which results in higher RNA yields. We suggested using the TURBO DNA-free™ kit (Thermo Fisher Scientific) for DNase treatment.
To define the desired input amount of RNA, laboratories were advised to quantify the RNA extracts using fluorescence or electrophoresis based quantification methods. The following methods were recommended: 1) Quant-iT RiboGreen RNA kit / Fluorescence microplate reader (high-and low-range protocol options) (Thermo Fisher Scientific); 2) QuantiFluor RNA System / QuantiFluor-ST Fluorometer (Promega, Madison, WI); 3) Quant-iT RNA assay kit / Qubit Fluorometer (Thermo Fisher Scientific); 4) Bioanalyzer (normally bad RNA quality / low RIN numbers with these kind of samples). In case the laboratories had no means of quantifying the RNA extracts, the organising laboratory proposed a specific input volume for each stain (Table S3).

Ingold S., Dørum G., Hanson E., Berti A., Branicki W., Brito P., Elsmore P., Gettings K.B., Giangasparo F., Gross T.E., Hansen S., Hanssen E.N., Kampmann M.L., Kayser M., Laurent F.X., Morling N., Mosquera-Miguel A., Parson W., Phillips C., Porto M.J., Pośpiech E., Roeder A.D., Schneider P.M., Schulze Johann K., Steffen C.R., Syndercombe-Court D., Trautmann M., van den Berge M., van der Gaag K.J., Vannier J., Verdoliva V., Vidaki A., Xavier C., Ballantyne J, & Haas C. (2018). Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise. Forensic science international. Genetics, 34.

+ Open protocol

+ Expand

RNA-Seq Workflow for Bacterial Transcriptomics

Check if the same lab product or an alternative is used in the 5 most similar protocols

The quality control of both the RNA quantity available and its quality was based on the QuantiFluor RNA System (Promega, Madison, WI, USA) and picoRNA chip on a bioanalyser 2100 (Agilent Technologies, Santa Clara, CA, USA), respectively. After quality control, two replicates remained available for the control and the starch-supplemented growth media while three replicates remained for each of the other growth conditions. Then, bacterial mRNAs were amplified by two rounds of in vitro transcription (IVT) using the ExpressArt Bacterial mRNA amplification kit (AmpTec, Hamburg, Germany). Finally, RNA libraries were generated using the ScriptSeq Complete kit for bacteria (Illumina Inc, San Diego, CA, USA) that allows performing directional RNA sequencing. RNA-sequencing was performed by a private company (Viroscan 3D, Lyon, France). The Appendix A details the protocols that are organized into three steps: 1) quality control, 2) total RNA amplification, and 3) library generations, respectively.

Sanhueza D., Guégan J.F., Jordan H, & Chevillon C. (2019). Environmental Variations in Mycobacterium ulcerans Transcriptome: Absence of Mycolactone Expression in Suboptimal Environments. Toxins, 11(3), 146.

+ Open protocol

+ Expand

Transcriptome Profiling of Brushing Samples

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted from each brushing sample according to the manufacturer’s instructions using the miRNA easy Mini Kit (QIAGEN, Hilden, Germany). Quantification was performed using the QuantiFluor RNA System (Promega, Madison, WI), and 50 ng of RNA was used as input to the TruSeq RNA Access Library Prep procedure (Illumina, San Diego, CA), which enriches for the coding transcriptome. Libraries meeting quality control criteria for amplification yields were sequenced using NextSeq 500 instruments (2 × 75 bp paired-end reads) with the High Output Kit (Illumina, San Diego, CA).
Raw sequencing (FASTQ) files were aligned to the Human Reference assembly 37 (Genome Reference Consortium) using the STAR RNA-seq aligner software [18 (link)]. Samples and sequencing runs that met pre-specified criteria for the number and quality of reads, as well as genomic representation and read depth, were used for downstream analysis. Based on annotated Ensembl genes, uniquely mapped reads were summarized using HTSeq [19 (link)]. The sequencing data was further filtered and normalized as described in [14 (link)].

Johnson M.K., Wu S., Pankratz D.G., Fedorowicz G., Anderson J., Ding J., Wong M., Cao M., Babiarz J., Lofaro L., Walsh P.S., Kennedy G.C, & Huang J. (2021). Analytical validation of the Percepta genomic sequencing classifier; an RNA next generation sequencing assay for the assessment of Lung Cancer risk of suspicious pulmonary nodules. BMC Cancer, 21, 400.

+ Open protocol

+ Expand

RNA Purification and Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols

After RNA isolation was performed using the different methods described above, all RNAs except for Trizol isolated, were further purified using an equal volume of chloroform (Fisher-Sci., #366919), centrifuged (16,000× g, 10 min, 4 °C), a back-extraction with TE buffer (10 mM Tris pH 7.6, 1 mM EDTA pH 8.0) to ensure no loss of RNA during extraction and then the aqueous phases containing RNA were precipitated with 100% ethanol in the presence of Glycogen and 0.6 M sodium acetate and incubated at −20 °C overnight. RNA samples were centrifuged (16,000× g, 1 h, 4 °C) and the supernatant removed. RNAs were washed with 80% ethanol, air dried and solubilized in DEPC treated dH₂O and then quantified using the QuantiFluor^® RNA System (Promega, #E3310) and Nanodrop 2000 (Thermo Scientific). RNA quality, like the RNA Integrity Number (RIN) and the DV200 was assessed using the Bioanalyzer 2100 with the corresponding 2100 Expert Software from Agilent.

Köhler S.A., Brandl L., Strissel P.L., Gloßner L., Ekici A.B., Angeloni M., Ferrazzi F., Bahlinger V., Hartmann A., Beckmann M.W., Eckstein M, & Strick R. (2022). Improved Bladder Tumor RNA Isolation from Archived Tissues Using Methylene Blue for Normalization, Multiplex RNA Hybridization, Sequencing and Subtyping. International Journal of Molecular Sciences, 23(18), 10267.

+ Open protocol

+ Expand

RNA-seq Library Preparation and Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

Library preparation and sequencing analysis were conducted at Bioengineering Lab. Co., Ltd. (Sagamihara, Japan). The concentration of the total RNA was measured using the Quantus Fluorometer with the QuantiFluor RNA system (Promega, Madison, WI, USA). The quality of the RNA was then analyzed using the 5200 Fragment Analyzer System (Agilent Technologies, Santa Clara, CA, USA). RNA sequencing libraries were prepared using the MGIEasy RNA Directional Library Prep Set (MGI Tech, Shenzhen, China) according to the manufacturer’s instructions. The concentration of the prepared library solution was determined using the Qubit 3.0 Fluorometer with the dsDNA HS Assay Kit (Thermo Fisher Scientific). The quality of the library was analyzed using the 5200 Fragment Analyzer System with the dsDNA 915 Reagent Kit (Agilent Technologies). Single-stranded circular DNA was prepared using the constructed library and the MGIEasy Circularization Kit (MGI Tech), and DNA nanoballs (DNBs) were prepared using the DNBSEQ-G400RS High-throughput Sequencing Kit (MGI Tech). The 200 bp paired-end sequencing was performed using the DNBSEQ-G400 (MGI Tech). The short-read data were deposited in the Read Archive of DDBJ (accession number DRA014441).

Megaly A.M., Miyashita M., Abdel-Wahab M., Nakagawa Y, & Miyagawa H. (2022). Molecular Diversity of Linear Peptides Revealed by Transcriptomic Analysis of the Venom Gland of the Spider Lycosa poonaensis. Toxins, 14(12), 854.

+ Open protocol

+ Expand

FFPE RNA Extraction and Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNA was extracted from 1 to 4 FFPE sections of 10 µm using Maxwell 16 LEV RNA FFPE extraction kit according to manufacturer's instructions. RNA samples were then quantified using QuantiFluor^® RNA System on a Quantus^™ Fluorometer (Promega).

Tachon G., Cortes U., Richard S., Martin S., Milin S., Evrard C., Lamour C, & Karayan‐Tapon L. (2019). Targeted RNA‐sequencing assays: a step forward compared to FISH and IHC techniques?. Cancer Medicine, 8(18), 7556-7566.

+ Open protocol

+ Expand

Single-cell RNA-seq of Mouse Neurons

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from flow-sorted neurons using Trizol LS (Invitrogen) and the RNA Clean and Concentrator-5 kit (Zymo Research). RNA concentration and quality was determined using Quantifluor RNA system (Promega) and Fragment analyzer (Advanced Analytics). Samples were converted to cDNA, depleted of rRNA transcripts, and amplified using the TRIO RNA-seq kit for mouse (Nugen Inc). Single-end sequencing was performed on a Nextseq 550 (Illumina) for 76 cycles. Read quality was assessed using FASTQC (https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and adapters were trimmed using Trimmomatic (Bolger et al., 2014 (link)). Filtered and trimmed reads were aligned to the mouse genome (mm10, UCSC) using Tophat2 (v2.1.1) (Kim et al., 2013 (link)).

Donovan L.J., Spencer W.C., Kitt M.M., Eastman B.A., Lobur K.J., Jiao K., Silver J, & Deneris E.S. (2019). Lmx1b is required at multiple stages to build expansive serotonergic axon architectures. eLife, 8, e48788.

+ Open protocol

+ Expand

Isolation of Fly Flight Muscle RNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

For each replicate, flight muscles from seven Mef2-GAL4, UAS-GFP-GMA pupae at 24 h or 32 h APF were dissected in ice-cold PBS treated with DEPC using a fluorescent binocular. Flight muscles were collected in an Eppendorf tube and centrifuged at 2000 g for 5 min. The flight muscle pellet was re-suspended in TRIzol™, shock-frozen in liquid nitrogen and kept at -80°C.
RNA was isolated directly from the TRIzol muscle samples using a 96 well plate extraction kit (Direct-zol™-96 RNA, Zymo Research, #R2054): after thawing to room temperature in 1,5 ml Eppendorf tubes, the tissue samples were homogenized using a small pestle, followed by nucleic acid precipitation with 100% ethanol. This suspension was then transferred to the 96-well plate containing the purification columns. DNA digestion was performed 'in column' according to the kit instructions. Total RNA was eluted with 25 μl of RNase-free water and was quantified using the Quantifluor RNA System (Promega, #E3310).

Kaya-Çopur A., Marchianò F., Hein M.Y., Alpern D., Russeil J., Luis N.M., Mann M., Deplancke B., Habermann B., & Schnorrer F. (2020). The Hippo pathway controls myofibril assembly and muscle fiber growth by regulating sarcomeric gene expression.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!