For purification of up to 100 µg total RNA from cells/tissues using gDNA Eliminator columns
- Unique gDNA Eliminator columns avoid the need for DNase
- Efficient removal of genomic DNA
- Highly reproducible yields of RNA in minutes
- High-performance RNA for sensitive applications
The RNeasy Plus Mini Kit integrates fast, convenient purification of up to 100 µg RNA with effective elimination of genomic DNA. Cell or tissue lysates are spun through gDNA Eliminator spin columns to remove genomic DNA. Total RNA is then purified using RNeasy Mini spin columns. The kit can be automated on the QIAcube Connect. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. For smaller samples, the RNeasy Plus Micro Kit (spin-column binding capacity of 45 µg RNA) is also available. Request a quote for a trial kit.
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製品名
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Cat. no.
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List price:
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RNeasy Plus Mini Kit (50)
For 50 minipreps: RNeasy Mini Spin Columns, gDNA Eliminator Spin Columns, Collection Tubes, RNase-Free Water and Buffers
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74134
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RNeasy Plus Mini Kit (250)
For 250 minipreps: RNeasy Mini Spin Columns, gDNA Eliminator Spin Columns, Collection Tubes, RNase-Free Water and Buffers
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74136
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RNeasy Plus Mini Kit 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。
High, reproducible RNA yields.|効率的な組織ゲノムDNAの除去|RNeasy Plus調製法|効率的な細胞ゲノムDNAの除去|アレイ解析が即可能なRNA|
Total RNA was purified in duplicate from different amounts of Jurkat cells using the RNeasy Plus Mini Kit or a similar kit from Supplier AV. Real-time RT-PCR assays for β-actin were performed (40 cycles). The lower CT values with the RNeasy Kit demonstrate greater RNA yields. With the kit from Supplier AV, no transcript was detectable in RNA purified from 102 cells. |RNeasy Plus Mini Kitあるいは他社のキットを用いて異なるマウス組織(各サンプル10 mg)からトータルRNAをduplicateで精製した。精製したRNA中のDNAの混入量を調べるためにc-jun用のリアルタイムPCRアッセイを行なった。|短い調製により、25分未満でRNA精製とゲノムDNAの除去が可能。|[A] RNeasy Plus Mini Kitあるいは[B] Supplier AVのゲノムDNA除去と組み合わせたRNA精製キットを用い、Jurkat細胞サンプル(サンプルあたり1 x 106個)からトータルRNAを精製した。β-actin用のリアルタイムRT-PCRアッセイを逆転写酵素あり(+RT)またはなし(-RT)でduplicateで行なった。-RT曲線は、RNeasy Plus Mini Kitを用いて精製したRNAに混在するゲノムDNAが事実上ないことを示す。|RNeasy Plus Mini Kitを用い、1 x 106個のHeLa細胞からトータルRNAをduplicateで精製した。cRNAは、GeneChip IVT Labeling Kitを用い、複製RNAサンプル(各3.5 μg)から調製した。cRNAサンプル(各15 μg)をGeneChip Human Genome U133Aプローブアレイで解析した。スキャッタープロットは2種類のサンプル間の相関性(ピアソンの相関係数[r]0.996)を示している。赤色:両方のサンプルに存在する遺伝子、青色:1サンプルに存在しないまたは最低限の遺伝子、黄色:両方のサンプルに存在しないまたは最低限の遺伝子。|
原理
Cells and easy-to-lyse tissues are lysed and homogenized in highly denaturing guanidine-isothiocyanate–containing Buffer RLT Plus, which immediately inactivates RNases to ensure isolation of intact RNA. The lysate is then passed through a gDNA Eliminator spin column. This column, in combination with the high-salt buffer, selectively and efficiently removes genomic DNA. Ethanol is added to provide appropriate binding conditions for RNA, and the sample is applied to an RNeasy MinElute spin column. These specialized columns contain a silica membrane that specifically binds RNA from lysed cells.
操作手順
Total RNA is purified from up to 107 cells or 30 mg tissue. A short workflow enables RNA purification with genomic DNA removal in less than 25 minutes (see flowchart "RNeasy Plus procedure"). Samples are first lysed and homogenized. The lysate is passed through a gDNA Eliminator spin column, ethanol is added to the flow-through, and the sample is applied to an RNeasy MinElute spin column. RNA binds to the membrane and contaminants are washed away. High quality RNA is eluted in 30 µl, or more, of water.
Different protocols are available for different starting materials. The protocols differ mainly in the lysis and homogenization of the sample. Once the sample is applied to the gDNA Eliminator spin column, the protocols are similar. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. The RNeasy Plus procedure can be modified to allow the purification of total RNA containing small RNAs, such as miRNA, from cultured cells.
When disrupting and homogenizing tissues in Buffer RLT Plus (supplied with the RNeasy Plus Mini Kit), excessive foaming may occur. This foaming is substantially reduced by adding Reagent DX (supplied separately) to Buffer RLT Plus before starting disruption and homogenization. Reagent DX has been carefully tested with the kit, and has no effect on RNA purity or on downstream applications.
アプリケーション
RNA purified using the RNeasy Plus Mini Kit 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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30-50 µl
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Format
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Spin column
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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, cells
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Processing
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Manual (centrifugation)
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Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
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Total RNA
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Sample amount
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30 mg (<700 µl)
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Technology
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Silica technology
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Time per run or per prep
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25 minutes
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Yield
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Varies
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FAQ ID -1024
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表示
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FAQ ID -1037
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FAQ ID -1043
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FAQ ID -1087
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FAQ ID -1164
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FAQ ID -1258
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FAQ ID -1577
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FAQ ID -1616
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FAQ ID -1619
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FAQ ID -1734
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FAQ ID -1996
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FAQ ID -2340
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FAQ ID -2794
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FAQ ID -2797
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FAQ ID - 3390
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FAQ ID -627
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FAQ ID -2793
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FAQ ID -728
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FAQ ID -2248
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FAQ ID -2796
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gDNA Eliminatorカラムを用いた動物細胞および溶解しやすい動物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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詳細を表示
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詳細を表示
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There are two protocols: follow Protocol 1 if you want to purify total RNA containing miRNA, or follow Protocol 2 if you want to purify small RNA (includes miRNA, 5S rRNA, and tRNA) and larger RNA (>200 nt) separately.
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詳細を表示
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Download Safety Data Sheets for QIAGEN product components.
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表示
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Poster for download
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詳細を表示
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画像
High, reproducible RNA yields.
Total RNA was purified in duplicate from different amounts of Jurkat cells using the RNeasy Plus Mini Kit or a similar kit from Supplier AV. Real-time RT-PCR assays for β-actin were performed (40 cycles). The lower CT values with the RNeasy Kit demonstrate greater RNA yields. With the kit from Supplier AV, no transcript was detectable in RNA purified from 102 cells.
効率的な組織ゲノムDNAの除去
RNeasy Plus Mini Kitあるいは他社のキットを用いて異なるマウス組織(各サンプル10 mg)からトータルRNAをduplicateで精製した。精製したRNA中のDNAの混入量を調べるためにc-jun用のリアルタイムPCRアッセイを行なった。
RNeasy Plus調製法
短い調製により、25分未満でRNA精製とゲノムDNAの除去が可能。
効率的な細胞ゲノムDNAの除去
[A] RNeasy Plus Mini Kitあるいは[B] Supplier AVのゲノムDNA除去と組み合わせたRNA精製キットを用い、Jurkat細胞サンプル(サンプルあたり1 x 106個)からトータルRNAを精製した。β-actin用のリアルタイムRT-PCRアッセイを逆転写酵素あり(+RT)またはなし(-RT)でduplicateで行なった。-RT曲線は、RNeasy Plus Mini Kitを用いて精製したRNAに混在するゲノムDNAが事実上ないことを示す。
アレイ解析が即可能なRNA
RNeasy Plus Mini Kitを用い、1 x 106個のHeLa細胞からトータルRNAをduplicateで精製した。cRNAは、GeneChip IVT Labeling Kitを用い、複製RNAサンプル(各3.5 μg)から調製した。cRNAサンプル(各15 μg)をGeneChip Human Genome U133Aプローブアレイで解析した。スキャッタープロットは2種類のサンプル間の相関性(ピアソンの相関係数[r]0.996)を示している。赤色:両方のサンプルに存在する遺伝子、青色:1サンプルに存在しないまたは最低限の遺伝子、黄色:両方のサンプルに存在しないまたは最低限の遺伝子。
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