FAQ-532
Is it possible to isolate both RNA and recombinant 6xHis-tagged protein from the same sample?
We have no experimental data for this application. However, buffer RLT of the RNeasy Kits for RNA isolation does not contain substances incompatible with Ni-NTA purification of His-tagged proteins. It should be possible to first extract RNA from a sample by following the RNeasy procedure, save the flow-through from the binding step as well as from the RW1 wash, and apply the combined fractions onto a Ni-NTA column for binding of His-tagged proteins. Follow our recommendations for purification of 6xHis-tagged proteins using Ni-NTA resins outlined in the QIAexpressionist handbook.
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RNeasy Kits
RNeasy Kits are the gold standard for total RNA isolation. They provide fast purification of high-quality RNA from small to large amounts of cells, tissues, and yeast using silica membrane RNeasy spin columns or 96-well plates. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor II or TissueLyser II or LT system. The RNeasy 96 Kit enables high-throughput purification of total RNA from up to 96 cultured cell samples using silica membrane RNeasy 96 plates. A dedicated RNeasy QIAcube Kit enables automated purification of 1–12 samples on the QIAcube Connect.
RNeasy Protect Kits
RNeasy Protect Mini Kits enable stabilization of RNA in tissue samples, RNA and DNA in sorted or cultured cells, RNA in human saliva samples and RNA in bacterial samples. The RNeasy Protect Mini Kit and Bacteria Mini Kit include RNeasy spin columns for purifying up to 100 µg of high-quality RNA using silica-membrane technology. The RNeasy Protect Cell Mini Kit provides the RNeasy Plus Mini Kit for purification of total RNA. The RNeasy Protect Saliva Mini Kit includes the RNeasy Micro Kit, which purifies and concentrates total RNA using specialized RNeasy MinElute spin columns. The RNeasy Protect Mini, RNeasy Protect Cell Mini and RNeasy Protect Bacteria Mini Kits can be automated on the QIAcube Connect.
RNeasy Plant Mini Kit
The RNeasy Plant Mini Kit includes QIAshredder spin columns for homogenizing and filtering viscous plant or fungal lysates, and RNeasy spin columns for purifying up to 100 μg of high-quality RNA using silica-membrane technology. Purification can be fully automated on the QIAcube Connect. The kit can also be used in combination with a TissueRuptor or TissueLyser system, which provide efficient disruption and homogenization of plant samples.
RNeasy Lipid Tissue Mini Kit
The RNeasy Lipid Tissue Mini Kit includes QIAzol Lysis Reagent for lysing fatty tissues and other types of tissue, and RNeasy spin columns for purifying up to 100 µg of high-quality RNA. The kit can be automated using the QIAcube Connect. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent (nonfatty tissues only) or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system.
RNeasy Fibrous Tissue Mini Kit
The RNeasy Fibrous Tissue Mini Kit includes proteinase K for removing abundant protein in fiber-rich tissue samples, and RNeasy spin columns for purifying up to 100 µg of high-quality RNA. The kit can be automated on the QIAcube Connect. Fibrous tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. For larger samples, the RNeasy Fibrous Tissue Midi Kit (spin-column binding capacity of 1 mg RNA) is also available.
Ni-NTA Superflow
Ni-NTA Superflow is an affinity chromatography matrix for purifying recombinant proteins carrying a His tag.
Histidine residues in the His tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions with high specificity and affinity. Cleared cell lysates are loaded onto the matrices. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, His-tagged proteins are eluted in buffer under native or denaturing conditions.
Ni-NTA Spin System
Ni-NTA silica combines Ni-NTA with a macroporous silica support material optimized to suppress nonspecific hydrophobic interactions. Ni-NTA Spin Columns (His-protein purification spin columns) in the Ni-NTA Spin Kit and available separately provide Ni-NTA silica in a convenient microspin format for easy preparation of multiple samples in parallel. They provide a simple method for functional screening of engineered proteins, selection of clones expressing full-length translation products and comparison of expression levels. Each spin column can purify up to 300 µg of His-tagged protein. Like all Ni-NTA matrices, Ni-NTA spin columns can be used for one-step protein purification under native or denaturing conditions. The Ni-NTA Spin Kit is a complete kit for spin purification of His-tagged proteins. It can be automated on the QIAcube Connect (see image "QIAcube Connect").
Ni-NTA Agarose
Ni-NTA Agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a His tag. Histidine residues in the His tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions with high specificity and affinity. Cleared cell lysates are loaded onto the matrices. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, His-tagged proteins are eluted in buffer under native or denaturing conditions