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Rna minelute kit

Manufactured by Qiagen
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The RNA Minelute kit is a laboratory tool designed for the rapid purification of RNA from various sample types. It utilizes a silica-based membrane technology to efficiently capture and purify RNA molecules, enabling their subsequent analysis and downstream applications.

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Lab products found in correlation

Rneasy kit, by Qiagen (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Turbo dnase, by Thermo Fisher Scientific (2 mentions) Bacterial rna protect buffer, by Qiagen (2 mentions) Facsaria 2, by BD (1 mentions) Papain, by Worthington (1 mentions) Lightcycler 480 instrument, by Roche (1 mentions) High capacity reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Sybr select master mix, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

MinElute kit (274 citations) RNeasy MinElute RNA Cleanup kit (5 citations) Rneasy MinElute RNA purification kit (4 citations) MinElute RNA cleanup kit (3 citations)

4 protocols using rna minelute kit

1

Dissociation and RNA-seq of Drosophila Pupal Brain

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For tissue dissociation, pupal brain tissue dissected at 40 hr after pupal formation was incubated with a Papain (Worthington) and Liberase TM protease cocktail (Roche) at 25°C for 15 min in a microfuge shaker at 1,000 rpm. At 5 and 10 min into this incubation, the tissue was pipetted up and down with a P200. At 15 min, the sample was passed through a 25G 5/8-gauge needle until the tissue was completely dissociated. Digestion was inactivated by the addition of rich media with serum, and the cell suspension was passed through a 70 µm filter. To concentrate the cells, the suspension was spun down at 1,600 rpm for 8 min at 4°C. After decanting the supernatant, cells were re-suspended in ~500 µl of rich media and sorted in a BD FACSAria II.
RNA was then isolated from sorted cells using the RNA-min elute kit from QIAGEN. mRNA was amplified in a linear fashion using Arcuturus RiboAmp HS kit (Life Technologies). cDNA was then generated for quality assessment and paired-end Illumina sequencing libraries were prepared.
Detailed protocols are available upon request.
Tan L., Zhang K.X., Pecot M.Y., Nagarkar-Jaiswal S., Lee P.T., Takemura S.Y., McEwen J.M., Nern A., Xu S., Tadros W., Chen Z., Zinn K., Bellen H.J., Morey M, & Zipursky S.L. (2015). Ig Superfamily Ligand and Receptor Pairs Expressed in Synaptic Partners in Drosophila. Cell, 163(7), 1756-1769.
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2

Quantitative PCR for Gene Expression

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qPCR was performed as described previously (Sur et al., 2012b (link)). Essentially, total RNA was isolated from whole tissue by homogenizing in RNA Bee reagent (ambios AMS Biotechnology) followed by RNA isolation using Qiagen’s RNA MinElute kit according to manufacturers' protocols. 0.5–1 µg of total RNA was reverse transcribed using high capacity reverse transcription kit in a 20 μl reaction (Applied Biosystems). Quantitative PCR in triplicates was performed using the SYBR select master mix (Applied Biosystems) on the LightCycler 480 instrument (Roche). For normalization, mouse β-actin transcripts were used as internal controls. Following primer pairs were used for quantitative PCR analysis.
Myc-Fw: 5'-GGGGCTTTGCCTCCGAGCCT-3', Myc-Rev: 5'-TGAGGGGCATC GTCGTGGCT-3', β-actin-Fw: 5'CTGTCGAGTCGCGTCCACCCG-3', β-actin-Rev: 5'-CATGCCGGAGCCGTTGTCGAC-3'.
Dave K., Sur I., Yan J., Zhang J., Kaasinen E., Zhong F., Blaas L., Li X., Kharazi S., Gustafsson C., De Paepe A., Månsson R, & Taipale J. (2017). Mice deficient of Myc super-enhancer region reveal differential control mechanism between normal and pathological growth. eLife, 6, e23382.
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3

Quantifying T6SS gene expression in P. aeruginosa

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Expression of T6SS effector and structural genes was measured in wild-type P. aeruginosa PAO1. Cultures were grown for 16 hrs in LB broth, then cells (5 μL) were spotted on 0.2 μm nitrocellulose filters placed upon agar plates. Agar plate composition and incubation conditions are the same as employed in PAEE (Supplementary Table 4). Spots were harvested from nitrocellulose and resuspended into Bacterial RNA Protect buffer (Qiagen). Cells were lysed by lysozyme treatment (1 mg/ml for 10 min) followed by sonication. RNA was purified using the RNeasy kit (Qiagen), residual DNA removed by Turbo DNAse (Invitrogen), and remaining RNA purified and concentrated using the RNA Minelute kit (Qiagen). cDNA was synthesized using the High Capacity cDNA Reverse Transcription Kit (Thermo Fisher, Watham, MA). Gene expression was measured using SYBR Green-based quantitative PCR with primers targeting each gene of interest, and normalized to the housekeeping gene rpoD. Primer sequences are available upon request. Two independent experiments with three technical replicates included were performed for each measurement.
LaCourse K.D., Peterson S.B., Kulasekara H.D., Radey M.C., Kim J, & Mougous J.D. (2018). Conditional toxicity and synergy drive diversity among antibacterial effectors. Nature microbiology, 3(4), 440-446.
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4

Quantifying T6SS gene expression in P. aeruginosa

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Expression of T6SS effector and structural genes was measured in wild-type P. aeruginosa PAO1. Cultures were grown for 16 hrs in LB broth, then cells (5 μL) were spotted on 0.2 μm nitrocellulose filters placed upon agar plates. Agar plate composition and incubation conditions are the same as employed in PAEE (Supplementary Table 4). Spots were harvested from nitrocellulose and resuspended into Bacterial RNA Protect buffer (Qiagen). Cells were lysed by lysozyme treatment (1 mg/ml for 10 min) followed by sonication. RNA was purified using the RNeasy kit (Qiagen), residual DNA removed by Turbo DNAse (Invitrogen), and remaining RNA purified and concentrated using the RNA Minelute kit (Qiagen). cDNA was synthesized using the High Capacity cDNA Reverse Transcription Kit (Thermo Fisher, Watham, MA). Gene expression was measured using SYBR Green-based quantitative PCR with primers targeting each gene of interest, and normalized to the housekeeping gene rpoD. Primer sequences are available upon request. Two independent experiments with three technical replicates included were performed for each measurement.
LaCourse K.D., Peterson S.B., Kulasekara H.D., Radey M.C., Kim J, & Mougous J.D. (2018). Conditional toxicity and synergy drive diversity among antibacterial effectors. Nature microbiology, 3(4), 440-446.
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