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Rneasy columns and reagents

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RNeasy columns and reagents are a set of products from Qiagen used for the isolation and purification of ribonucleic acid (RNA) from various biological samples. The core function of these products is to facilitate the extraction and concentration of RNA molecules, which are essential for various downstream applications in molecular biology, genomics, and transcriptomics.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (3 mentions) High capacity rna cdna kit, by Thermo Fisher Scientific (2 mentions) Taqman primer sets, by Thermo Fisher Scientific (1 mentions)

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RNeasy columns (885 citations) RNeasy reagents (26 citations)

3 protocols using rneasy columns and reagents

1

Brain RNA Extraction and qPCR Analysis

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Brain tissue was homogenized directly in Trizol (Invitrogen) using a motorized grinder. Chloroform (1:5) was added, samples were agitated then centrifuged at 13,000g for 15 min at 4 degrees, and the chloroform layer was removed, diluted 1:1 in 70% ethanol, then purified using RNeasy columns and reagents (Qiagen, Valencia, CA). Reverse transcription was performed using high‐capacity RNA‐cDNA kit (Applied Biosystems (ABI), Carlsbad, CA) with 1 ug RNA per 20 uL reaction. Real‐time qPCR was performed using ABI Taqman primers and reagents on an ABI Prizm 7500 thermocycler according to manufacturer's instructions. All mRNA measurements were normalized to beta‐actin (Actb) mRNA levels. Taqman primer sets were obtained from Life Technologies using their proprietary sequences.
Musiek E.S., Xiong D.D., Patel T., Sasaki Y., Wang Y., Bauer A.Q., Singh R., Finn S.L., Culver J.P., Milbrandt J, & Holtzman D.M. (2016). Nmnat1 protects neuronal function without altering phospho‐tau pathology in a mouse model of tauopathy. Annals of Clinical and Translational Neurology, 3(6), 434-442.
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2

RNA Isolation and qPCR Analysis of Mouse Brain

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RNA isolation from mouse brain was performed as previously described (Musiek et al., 2013 (link)). Briefly, anterior cortex was homogenized by trituration through a 23-gauge needle in TRIzol (Invitrogen). Chloroform (1:5) was added then samples were mixed, and centrifuged (13,000 g; 15 min; 4 °C). Chloroform was removed, and samples were diluted 1:1 in 70% ethanol and purified using RNeasy columns and reagents (QIAGEN). RNA concentration was measured using a NanoDrop spectrophotometer. Reverse transcription was performed using a high-capacity RNA-cDNA kit (Applied Biosystems [ABI]) with 1 μg RNA per 20 μL reaction. Real-time qPCR was performed with ABI TaqMan primers and reagents on an ABI Prizm 7500 thermocycler according to the manufacturer’s instructions. Primers used: Grin2a (TaqMan; Mm00433802_m1), Grin2b (TaqMan; Mm00433820_m1), Gabra5 (IDT; Mm.PT.58.5845925), Actb (TaqMan; Mm01205647_g1). All mRNA measurements were normalized to Actb (β-actin) then to wildtype + H2O group mRNA levels.
Day S.M., Gironda S.C., Clarke C.W., Snipes J.A., Nicol N.I., Kamran H., Vaughan W., Weiner J.L, & Macauley S.L. (2022). Ethanol exposure alters Alzheimer’s-related pathology, behavior, and metabolism in APP/PS1 mice. Neurobiology of disease, 177, 105967.
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3

RNA Isolation and qPCR Analysis of Mouse Brain

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNA isolation from mouse brain was performed as previously described (Musiek et al., 2013 (link)). Briefly, anterior cortex was homogenized by trituration through a 23-gauge needle in TRIzol (Invitrogen). Chloroform (1:5) was added then samples were mixed, and centrifuged (13,000 g; 15 min; 4 °C). Chloroform was removed, and samples were diluted 1:1 in 70% ethanol and purified using RNeasy columns and reagents (QIAGEN). RNA concentration was measured using a NanoDrop spectrophotometer. Reverse transcription was performed using a high-capacity RNA-cDNA kit (Applied Biosystems [ABI]) with 1 μg RNA per 20 μL reaction. Real-time qPCR was performed with ABI TaqMan primers and reagents on an ABI Prizm 7500 thermocycler according to the manufacturer’s instructions. Primers used: Grin2a (TaqMan; Mm00433802_m1), Grin2b (TaqMan; Mm00433820_m1), Gabra5 (IDT; Mm.PT.58.5845925), Actb (TaqMan; Mm01205647_g1). All mRNA measurements were normalized to Actb (β-actin) then to wildtype + H2O group mRNA levels.
Day S.M., Gironda S.C., Clarke C.W., Snipes J.A., Nicol N.I., Kamran H., Vaughan W., Weiner J.L, & Macauley S.L. (2022). Ethanol exposure alters Alzheimer’s-related pathology, behavior, and metabolism in APP/PS1 mice. Neurobiology of disease, 177, 105967.
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