Rneasy mini kit

Manufactured by Bio-Rad

Sourced in United States, Germany

The RNeasy Mini Kit is a laboratory product designed to isolate and purify RNA from various biological samples. It utilizes a silica-membrane-based technology to efficiently capture and extract RNA, making it a versatile tool for researchers working with RNA analysis and manipulation.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

RNeasy Plus Mini Kit (3 citations) RNeasy kit (17 citations)

22 protocols using rneasy mini kit

Serum-induced Gene Expression Analysis

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

Gene expressions were induced in all bladder cell lines by culturing in 10% serum and another batch following a 24-h duration of serum starvation prior RNA purification (RNeasy Mini Kit, 74106), followed by converting 1 μg of samples to synthesized cDNA using a thermal cycler (C1000 Touch, Bio Rad) (High Capacity cDNA Reverse Transcription Kit, 4368813, Applied Biosystems).

Wong J.P., Wei R., Lyu P., Tong O.L., Zhang S.D., Wen Q., Yuen H.F., El-Tanani M, & Kwok H.F. (2017). Clinical and in vitro analysis of Osteopontin as a prognostic indicator and unveil its potential downstream targets in bladder cancer. International Journal of Biological Sciences, 13(11), 1373-1386.

+ Open protocol

+ Expand

BCR-ABL1 Kinase Domain Sequencing

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

RNA obtained from primary Ph⁺ leukemia cell lysates (QIAGEN RNeasy Mini Kit) served as template for cDNA synthesis (BioRad iScript cDNA Synthesis Kit) as recommended by the manufacturer. Amplification of the BCR-ABL1 kinase domain was done by two-step PCR to exclude amplification of normal ABL1 (Khorashad et al., 2006 (link)). PCR products were electrophoresed on a 2% agarose gel to confirm amplification, purified (QIAquick PCR Purification Kit; QIAGEN) and subjected to 1) conventional Sanger sequencing in both directions using BigDye terminator chemistry on an ABI3730 instrument (Khorashad et al., 2006 (link)), and 2) cloning and sequencing of amplified fragments introduced into E. coli TOP10 cells (TOPO cloning system; Invitrogen) (Khorashad et al., 2013 (link)). For cloning and sequencing, individual bacterial colonies (average: 85/specimen; range: 23-100), each carrying a recombinant plasmid with a single BCR-ABL1 kinase domain amplicon inserted, were subjected to BCR-ABL1 kinase domain amplification and Sanger sequenced in both directions (Beckman Coulter Genomics). DNA sequence analysis was done with Mutation Surveyor software (SoftGenetics) (O'Hare et al., 2009 (link)).

Zabriskie M.S., Eide C.A., Tantravahi S.K., Vellore N.A., Estrada J., Nicolini F.E., Khoury H.J., Larson R.A., Konopleva M., Cortes J.E., Kantarjian H., Jabbour E.J., Kornblau S.M., Lipton J.H., Rea D., Stenke L., Barbany G., Lange T., Hernández-Boluda J.C., Ossenkoppele G.J., Press R.D., Chuah C., Goldberg S.L., Wetzler M., Mahon F.X., Etienne G., Baccarani M., Soverini S., Rosti G., Rousselot P., Friedman R., Deininger M., Reynolds K.R., Heaton W.L., Eiring A.M., Pomicter A.D., Khorashad J.S., Kelley T.W., Baron R., Druker B.J., Deininger M.W, & O'Hare T. (2014). BCR-ABL1 Compound Mutations Combining Key Kinase Domain Positions Confer Clinical Resistance to Ponatinib in Ph Chromosome-Positive Leukemia. Cancer cell, 26(3), 428-442.

+ Open protocol

+ Expand

Quantification of Metabolic Gene Expression

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 J774A.1 cells for each group in triplicate using Qiagen's RNeasy Mini Kit and was reverse transcribed using iScript cDNA Synthesis Kit (BioRad Laboratories, Hercules, CA, USA). Polymerase chain reaction was performed using Piko-Real (Thermofisher, Fremont CA, USA) for RNA expression levels of genes of interest, namely, lactate dehydrogenase A and B isoforms (LDHA/B) and monocarboxylate transporters 1 and 4 isoforms (MCT1/4), using Taqman probes (Applied Biosystems, Foster City, CA, USA) as described previously 11 (link). β-actin was used as the housekeeping gene, and the relative difference from the control cells was calculated for each primer/probe combination.

Sriram R., Nguyen J., Santos J.D., Nguyen L., Sun J., Vigneron S., Van Criekinge M., Kurhanewicz J, & MacKenzie J.D. (2018). Molecular detection of inflammation in cell models using hyperpolarized 13C-pyruvate. Theranostics, 8(12), 3400-3407.

+ Open protocol

+ Expand

Quantitative RT-PCR for Transcriptome Analysis

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

Total RNA was extracted from the cells using RNeasy Mini Kit (cat# 74134) and 1 μg RNA used for cDNA synthesis (iScript cDNA Synthesis kit; cat# 1708891, Bio‐Rad). SYBR Green master mix (cat# 4309155, ThermoFisher Scientific) was used to determine mRNA expression according to the manufacturer's protocol using a QuantStudio 3 (Applied Biosystems). The RT‐PCR cycling conditions were as follows: DNA polymerase activation at 95°C for 10 min, followed by 40 PCR cycles, each cycle consisting of 95°C for 15 s (denature) and 60°C for 1 min (anneal/extend). GAPDH was used as a housekeeping control and data were analyzed with Applied Biosystems® qPCR analysis software (ThermoFisher Scientific).

Rajendran N.K., Liu W., Cahill P.A, & Redmond E.M. (2023). Caveolin‐1 inhibition mediates the opposing effects of alcohol on γ‐secretase activity in arterial endothelial and smooth muscle cells. Physiological Reports, 11(1), e15544.

+ Open protocol

+ Expand

Osteogenic Differentiation of hASCs

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

hASCs were seeded on a 12-well plate at a density of 2×10⁵ cells/well and incubated overnight. Cells were cultured for 14 days in osteogenic differentiation medium including test solution, and the medium was exchanged twice with a fresh one after 5 and 10 days. Then, total RNA was isolated with an RNeasy minikit (Bio-Rad) according to the manufacturer’s protocol. Then, cDNA was synthesized from extracted RNA using an iScript advanced cDNA-synthesis kit (Bio-Rad). All primers used were produced by DNA Technology (Aarhus, Denmark). The total volume of each sample was 20 μL, and samples were analyzed in duplicate with a MyCycler real-time PCR system (Bio-Rad). The thermocycling program consisted of an initial step of 3 minutes at 95°C and 40 cycles of 15 seconds at 95°C and an annealing/extension temperature of 60°C for 30 seconds. Sequences of the primer sets were:
Osteocalcin (OCN) forward 5ʹ-GAG CCC CAG TCC CCT ACC C-3ʹ, reverse 5ʹ-GCC TCC TGA AAG CCG ATG TG-3ʹ
Osteopontin (OPN) forward 5ʹ-TGA TGG CCG AGG TGA TAG TGT GGT-3ʹ, reverse 5ʹ-CCT GGG CAA CGG GGA TGG-3ʹ
Peptidylprolyl isomerase A (PPIA) forward 5ʹ-TCC TGG CAT CTT GTC CAT G-3ʹ, reverse 5ʹ-CCA TCC AAC CAC TCA GTC TTG-3ʹ.
Fold-change gene-expression levels were recorded using the ΔΔCt method, in which the data were normalized with the expression level of PPIA.

Jahed V., Vasheghani-Farahani E., Bagheri F., Zarrabi A., Fink T, & Lambertsen Larsen K. (2019). Enhanced Cellular Uptake Of Phenamil Through Inclusion Complex With Histidine Functionalized β-Cyclodextrin As Penetrative Osteoinductive Agent. International Journal of Nanomedicine, 14, 8221-8234.

+ Open protocol

+ Expand

Quantifying Immune Gene Expression

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

RNA from 5x10^6 splenocytes was extracted with the RNeasy Mini Kit (BioRad) and converted to cDNA using the Maxima First Strand cDNA Synthesis Kit (ThermoFisher Scientific). Quantitative reverse transcriptase polymerase chain reation (RT-qPCR) using iTaq Universal Syber Green Supermix (BioRad) and respective primers (Supplementary Table 1) was performed on cDNA to quantify the expression of Ifih1, Mx1, Ifit1, Hprt, and Oas1 genes. The plates were run on a Roche LightCycler 480 II. For ΔC_t values, gene expression was normalized to Hprt. Fold differences in mRNA expression (relative expression) were determined using the equation [2^(-ΔΔCt)].

Stock A.J., Gonzalez Paredes P., de Almeida L.P., Kosanke S.D., Chetlur S., Budde H., Wakenight P., Zwingman T.A., Rosen A.B., Allenspach E.J., Millen K.J., Buckner J.H., Rawlings D.J, & Gorman J.A. (2024). The IFIH1-A946T risk variant promotes diabetes in a sex-dependent manner. Frontiers in Immunology, 15, 1349601.

+ Open protocol

+ Expand

Splenocyte Stimulation and Gene Expression

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

Mice spleens were harvested immediately after mice were euthanized. Splenocytes were isolated by grinding moistened spleen sections between the ends of frosted microscope slides. The resultant cell suspensions were filtered through a 70-µm cell strainer (Thermo Fisher Scientific) and then counted, plated in equal numbers for all mice, and stimulated with 20 ng/ml mouse TNF for 24 h. mRNA was extracted using RNeasy Mini kit and was reverse-transcribed with the iScript cDNA synthesis kit (Bio-Rad Laboratories). The expression of the mediators Il6 (Mm00446190_m1), Tnfaip3 (Mm00437121_m1), and Traf1 (Mm00493827_m1) was measured using qPCR in a TaqMan Gene Expression Assay. The expression of these genes was analyzed using the 2^-ΔΔCT method in comparison to the housekeeping gene GUSB (Mm01197698_m1).

Badran Y.R., Dedeoglu F., Leyva Castillo J.M., Bainter W., Ohsumi T.K., Bousvaros A., Goldsmith J.D., Geha R.S, & Chou J. (2017). Human RELA haploinsufficiency results in autosomal-dominant chronic mucocutaneous ulceration. The Journal of Experimental Medicine, 214(7), 1937-1947.

+ Open protocol

+ Expand

Quantifying Gene Expression in Lymphoblastoid Cells

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

Total RNA was extracted from patient-derived lymphoblastoid cells using RNeasy mini kit and converted into cDNA by iScript cDNA synthesis kit (BioRad) according to manufacturer’s recommendations. qRT-PCR analysis was performed using Taqman probes for DNMT1-Hs00154749 and GAPDH-Hs02758991_g1 (Applied Biosystems) on Realplex Mastercycler (Eppendorf). Expression relative to GAPDH was calculated using 2^ΔΔCt

Pathak S., Miller J., Morris E.C., Stewart W.C, & Greenberg D.A. (2018). DNA methylation of the BRD2 promoter is associated with Juvenile Myoclonic Epilepsy in Caucasians. Epilepsia, 59(5), 1011-1019.

+ Open protocol

+ Expand

Heterologous Expression of Cycloartenol Synthase

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

To screen for potential OSCs, we performed a TBLASTX search in the transcriptome assembly of L. dendroidea [38 (link)] using the nucleotide sequence of the characterized A. annua cycloartenol synthase (CAS) (GenBank accession KM670093 [39 (link)]) as query. The domain composition of the sequence coding for the OSC from L. dendroidea was obtained through search for conserved domains using the National Center for Biotechnology Information (NCBI) Conserved Domain Database (CDD) [40 (link)].
Afterwards, L. dendroidea specimens obtained from an unialgal culture (see 38 for details) were frozen in liquid nitrogen and ground to a fine powder using a mortar and pestle. Total RNA was extracted with the Qiagen RNeasy^® Mini Kit (Hilden, Germany) and cDNA was prepared with the Bio-Rad iScript^TM cDNA Synthesis Kit (Hercules, United States). The full-length coding sequence of LdCAS was amplified from this L. dendroidea cDNA using the primers LdCAS_Fw and LdCAS_RV (Table 1). The obtained PCR fragment was Gateway^TM recombined into the Gateway^TM vector pDONR207 and the resulting entry clone was sequence verified and further recombined into the in-house generated destination vector pESC-URA-tHMG1-DEST [41 (link)] to yield pESC-URA-tHMG1-DEST[GAL1/LdCAS].

Calegario G., Pollier J., Arendt P., de Oliveira L.S., Thompson C., Soares A.R., Pereira R.C., Goossens A, & Thompson F.L. (2016). Cloning and Functional Characterization of Cycloartenol Synthase from the Red Seaweed Laurencia dendroidea. PLoS ONE, 11(11), e0165954.

+ Open protocol

+ Expand

Genotyping and Quantifying ClC-7 Expression

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

Genotyping of ClC-7 KO U2OS clone was performed using an Extract PCR-Kit (Bioline) with the following primers: GAGAACAAACACGGGGGCA; GCGTTCCCGAGTCCACC. Isolated total RNA from WT and KO U2OS cells (QIAGEN RNeasy Mini Kit) reverse-transcribed into cDNA (BIORAD iScript cDNA Synthesis Kit) and primers (indicated as followed) were used to quantify mRNA expression levels by real-time PCR using iQ SYBR Green Supermix (Bio-Rad) and CFX96 Touch Real-Time PCR Detection System (Bio-Rad). Primers used: GAPDH: 5′-GTCTCCTCTGACTTCAACAGCG-3′; 5′-ACCACCCTGTTGCTGTAGCCAA-3′. ClC-7: 5′-CCCACTCCAGCTCTTCTGTG-3′; 5′-ATAGGAGCCTGGTGGGTCATG-3′. Relative expression of ClC-7 mRNA was calculated by normalizing the expression values of WT and KO to those of GAPDH.

Leray X., Hilton J.K., Nwangwu K., Becerril A., Mikusevic V., Fitzgerald G., Amin A., Weston M.R, & Mindell J.A. (2022). Tonic inhibition of the chloride/proton antiporter ClC-7 by PI(3,5)P2 is crucial for lysosomal pH maintenance. eLife, 11, e74136.

+ 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!