RNeasy Plus Universal Kits
For purification of total RNA from all types of tissue using gDNA Eliminator Solution
- One-for-all tissue solution
- Time-saving integration of RNeasy and QIAzol technologies
- Fast non-enzymatic removal of genomic DNA
- Pure RNA for use in downstream applications
RNeasy Plus Universal Kits integrate fast, convenient RNA purification with effective elimination of genomic DNA. Optimized protocols enable purification of high-quality RNA from any type of tissue, even difficult-to-lyse tissues. The kits include QIAzol Lysis Reagent for lysing fatty tissue and other types of tissue and RNeasy spin columns for purifying high-quality RNA. Kits are available in two formats: the RNeasy Plus Universal Mini Kit enables purification of RNA from up to 50 mg tissue and the RNeasy Plus Universal Midi Kit enables purification of RNA from up to 250 mg tissue. The RNeasy Plus Universal Mini Kit can be automated on the QIAcube Connect.
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RNeasy Plus Universal Mini Kit (50)
For 50 RNA minipreps: RNeasy Mini Spin Columns, gDNA Eliminator Solution, Collection Tubes, RNase-Free Water and Buffers
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73404
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RNeasy Plus Universal Midi Kit (10)
For 10 RNA midipreps: RNeasy Midi Spin Columns, gDNA Eliminator Solution, Collection Tubes, RNase-Free Water and Buffers
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73442
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RNeasy Plus Universal Kits は分子生物学実験用です。疾病の診断、治療または予防の目的には使用することはできません。
ゲノムDNA の効率的な除去|高品質RNA|RNeasy Plus Universal MiniおよびMidi 操作手順|
RNAlaterで安定した表記量のラット腎臓から、RNeasy Plus Universal Mini Kitを用いてRNA を精製、または他社(AII 社)の沈殿試薬を用いて精製した。RNA溶出液を逆転写反応ありとなしで、QuantiFast Probe RT-PCR KitおよびRotor-Gene Qを用いてリアルタイムRT-PCRで解析した。|RNeasy Plus Universal Mini Kit を用いてgDNA Eliminator Solution を添加あるいは未添加という条件でラット組織からRNA を精製した。[A] 25 mgの凍結肝臓組織から精製したRNA を用いてリアルタイムRT-PCR を行なった。全サンプルのCT 値を比較した。gDNA Eliminator Solution を使用しなかった場合は-RT control でDNA の混入が検出されたが、gDNA Eliminator Solution を使用した場合はDNA の検出は微量、あるいは皆無であった。[B] 安定化した心臓組織、25 mg からRNA を精製した。QIAxcel システムを用いて得られたデータは、gDNA Eliminator Solution を使用してRNAを精製したかどうかに関わらず、RNA分解のないことを示している。||
パフォーマンス
The RNeasy Plus Universal procedure allows high, reproducible RNA yields from any type of tissue, including fatty tissues and difficult-to-lyse tissue (see table). Yields can vary due to factors such as species and developmental stage, especially with adipose tissues.
| Adipose tissue |
0.5–2.5 |
| Brain |
5–20 |
| Heart |
5–25 |
| Intestine |
10–60 |
| Kidney |
5–40 |
| Liver |
15–80 |
| Lung |
5–15 |
| Muscle |
5–35 |
| Skin |
2–5 |
| Spleen |
15–100 | Analysis of purified RNA by real-time RT-PCR, and on the QIAxcel system, demonstrates high-quality RNA with good integrity (see figure " High-quality RNA"). gDNA Eliminator Solution and RNeasy technologies efficiently remove most of the genomic DNA contamination without DNase treatment (see figure " Effective removal of genomic DNA").
原理
The RNeasy Plus Universal procedure enables purification of RNA from any type of tissue, including fatty or difficult-to-lyse tissues. RNeasy Plus Universal Kits integrate phenol/guanidine-based sample lysis and silica-membrane purification of total RNA. QIAzol Lysis Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of all kinds of tissues and to inhibit RNases. The highly efficient lysis and the subsequent removal of contaminants by organic phase extraction enable the use of larger amounts of tissue with RNeasy spin columns. gDNA Eliminator solution effectively removes genomic DNA contamination and significantly improves separation of DNA into the interphase.
操作手順
The RNeasy Plus Universal procedure is easy to follow and, compared to phenol/chloroform treatment with a subsequent cleanup procedure, much faster and more efficient with better RNA purity and yield (see flowchart "RNeasy Plus Universal Mini and Midi procedures"). Total RNA is purified from up to 50 mg tissue using the RNeasy Plus Universal Mini Kit or up to 250 mg tissue using the RNeasy Plus Midi Kit.
Tissue samples are homogenized in QIAzol Lysis Reagent using the TissueRuptor, TissueLyser LT, or TissueLyser II. After addition of gDNA Eliminator Solution and chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase with RNA is collected, mixed with ethanol, and RNA is purified using RNeasy spin columns. Total RNA binds to the spin column membrane, and phenol and other contaminants are efficiently washed away. High-quality RNA is eluted in RNase-free water.
The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. The RNeasy Plus Universal procedure can be modified to allow the purification of total RNA containing small RNAs, such as miRNA, from cultured cells.
アプリケーション
RNA purified using the RNeasy Plus Universal Kits is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as quantitative real-time RT-PCR. The purified RNA can also be used in other applications.
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Feature
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Specifications
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Applications
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Northern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
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Elution volume
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Mini: 30µl; Midi:150µl
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For automated processing
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Mini format can be automated on the QIAcube
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Format
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Mini and Midi Format
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Integrated removal of genomic DNA
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Yes
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Main sample type
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Tissue samples including difficult-to-lyse and demanding tissues
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Processing
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Manual (centrifugation), automated (QIAcube)
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Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
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RNA
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Sample amount
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Mini: 5-50 mg; Midi: 50-250 mg
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Technology
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QIAzol/Spin column technology
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FAQ ID -2338
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FAQ ID -2339
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FAQ ID -2793
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FAQ ID -2794
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FAQ ID -2796
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FAQ ID -2799
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FAQ ID - 3390
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FAQ ID -627
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FAQ ID -728
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FAQ ID -2797
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FAQ ID -2798
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FAQ ID -1024
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FAQ ID - 3388
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For purification of total RNA from all types of tissue
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詳細を表示
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RNeasy Plus Universal Mini Kit
RNeasy Plus Universal Midi Kit
すべてのタイプの組織からのトータルRNA 精製
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詳細を表示
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画像
ゲノムDNA の効率的な除去
RNAlaterで安定した表記量のラット腎臓から、RNeasy Plus Universal Mini Kitを用いてRNA を精製、または他社(AII 社)の沈殿試薬を用いて精製した。RNA溶出液を逆転写反応ありとなしで、QuantiFast Probe RT-PCR KitおよびRotor-Gene Qを用いてリアルタイムRT-PCRで解析した。
高品質RNA
RNeasy Plus Universal Mini Kit を用いてgDNA Eliminator Solution を添加あるいは未添加という条件でラット組織からRNA を精製した。[A] 25 mgの凍結肝臓組織から精製したRNA を用いてリアルタイムRT-PCR を行なった。全サンプルのCT 値を比較した。gDNA Eliminator Solution を使用しなかった場合は-RT control でDNA の混入が検出されたが、gDNA Eliminator Solution を使用した場合はDNA の検出は微量、あるいは皆無であった。[B] 安定化した心臓組織、25 mg からRNA を精製した。QIAxcel システムを用いて得られたデータは、gDNA Eliminator Solution を使用してRNAを精製したかどうかに関わらず、RNA分解のないことを示している。
RNeasy Plus Universal MiniおよびMidi 操作手順
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