RNeasy Midi Kit

細胞、組織、酵母からの最大1 mgまでのトータルRNA精製用
  • 迅速な操作により短時間で高品質なトータルRNAを実現
  • あらゆるダウンストリームアプリケーションに即使用可能なRNA
  • 中程度のスタートサンプル量で一定したRNA収量
  • フェノール/クロロホルム抽出不要
  • CsCl 勾配遠心分離、LiCl やエタノール沈殿は不要
RNeasy Midi Kitはシリカゲルメンブレンを用いたRNeasy Spin Column(RNA結合容量は1 mg)で細胞、組織、酵母から高品質RNAを迅速に精製します。組織サンプルはRNAlater RNA Stabilization ReagentまたはAllprotect Tissue Reagentで簡便に安定化され、TissueRuptorあるいはTissueLyserシステムで効率的に破砕されます。より少量あるいは多量のサンプルには、RNeasy Micro Kit(スピンカラム結合容量はRNA最高45 µg)、RNeasy Mini Kit(スピンカラム結合容量はRNA 100 µg)、RNeasy Maxi Kit(スピンカラム結合容量はRNA 6 mg)をご使用ください。
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RNeasy Midi Kit (50)
50 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers
75144
737,00 €
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The RNeasy Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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RNeasy Midi procedure.|High-quality RNA from a variety of samples.|RNeasy Midi spin column.|
Total RNA purified with the RNeasy Midi Kit is of high quality and is suitable for many downstream applications. Protocols are also included for cleanup of partially purified RNA, in vitro transcripts, and RNA from enzymatic reactions. An additional buffer is required (composition provided) for isolation of cytoplasmic RNA from eukaryotic cells. Lyticase, zymolase, or glass beads (required for yeast samples) are not provided. Amounts of RNA isolated from samples can vary due to the developmental stage, species, and growth conditions of the sample source. Since the RNeasy procedure enriches for RNA species >200 nt, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight RNAs.|Formaldehyde agarose gel and northern blot of total RNA purified with the RNeasy Maxi Kit. Total RNA (10 µg) isolated from each source was loaded per lane. All tissues were from mouse. Yeast: Saccharomyces cerevisiae; E. coli strain: HB101. 32P-labeled probes recognized (G) GAPDH; (E) translation elongation factor EF-1α; and (O) outer membrane protein A sequences. (E and O were kindly provided by P. Philippsen, University of Basel, Switzerland and U. Henning, Max Planck Institute of Biology, Tübingen, Germany, respectively.) B. subtilis was not probed. M: 0.24–9.5 kb RNA ladder. 7.5 kb band (indicated) in embryo, Huh7, and HeLa cell lanes is a nuclear precursor RNA.|The RNeasy Midi spin column contained in the RNeasy Midi Kit.|
Performance

RNeasy Midi Kitを用いて精製したトータルRNAは高品質で、多くのダウンストリームプリケーションに適しています(図"各種サンプルから得られた高品質RNA")。RNeasy Midi Kitを用いることにより、動物またはヒト細胞、動物またはヒト組織、酵母から簡単にトータルRNAを精製できます(表参照)。

RNeasy Midi Kitを用いて得られたトータルRNA収量
由来 スタートサンプル 平均収量(µg)

動物細胞
LMH
HeLa
COS-7
リンパ球(未刺激)


7 x 107
7 x 107
3 x 107
1 x 108

850
1000
950
50

マウス組織
肝臓

脾臓


200 mg
100 mg
200 mg

700
130
600

酵母細胞
S. cerevisiae


2 x 108

450
 

Principle

RNeasy Midi Kitは、中程度のサンプル量に対応し、効率的にトータルRNAを精製します。グアニジンイソチオシアネート溶解と、シリカゲルメンブレンによる迅速な精製を組み合わせたRNeasyテクノロジーにより、トータルRNA精製を簡便に行なうことができます。RNeasy Kitで精製した高品質のRNAはほとんどDNAを含みません。

Procedure
RNeasy Midi Kitを用いて、5 x 106~1 x 108の動物またはヒト細胞、20~250 mgの動物またはヒト組織、2 x 107~5 x 108の酵母から、簡単にトータルRNAを精製できます。まずサンプルを溶解し、その後ホモジナイズします。ライセートにエタノールを添加して最適な結合条件にします。その後ライセートをRNeasyシリカゲルメンブレンにアプライします(図"RNeasy Midi Spin Column")。RNA(最高1 mg容量)はメンブレンに結合し、全ての夾雑物は効率的に洗い流されます。微量のDNAに敏感なアプリケーションを行なう場合、カラム上での簡便なDNase処理によって残存するDNAを除去できます。濃縮された高純度のRNAは300~500 µlの水で溶出されます(図"RNeasy Midi操作手順")。各種のアプリケーションプロトコールも揃えています。
Applications

RNeasyテクノロジーで精製したRNAのA260/280比は、1.9~2.1(10 mM Tris·Cl、pH 7.5)で、以下を含む幅広いアプリケーションに最適です。

ノーザン、ドットおよびスロットブロット
エンドポイントRT-PCR
リアルタイム定量RT-PCR
アレイ解析
Poly A+RNA調製
Feature
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 300–500 µl
Format Spin column
Main sample type Tissue, cells
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 20–250 mg
Technology Silica technology
Time per run or per prep <1 hour
Yield Varies

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Kit Handbooks
1

For total RNA isolation from animal cells, animal tissues, bacteria, yeast, whole blood, and for RNA cleanup


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RNeasy Midi Procedure
RNeasy Midi procedure.

Total RNA purified with the RNeasy Midi Kit is of high quality and is suitable for many downstream applications. Protocols are also included for cleanup of partially purified RNA, in vitro transcripts, and RNA from enzymatic reactions. An additional buffer is required (composition provided) for isolation of cytoplasmic RNA from eukaryotic cells. Lyticase, zymolase, or glass beads (required for yeast samples) are not provided. Amounts of RNA isolated from samples can vary due to the developmental stage, species, and growth conditions of the sample source. Since the RNeasy procedure enriches for RNA species >200 nt, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight RNAs.

High-quality RNA from a variety of samples.
High-quality RNA from a variety of samples.
Formaldehyde agarose gel and northern blot of total RNA purified with the RNeasy Maxi Kit. Total RNA (10 µg) isolated from each source was loaded per lane. All tissues were from mouse. Yeast: Saccharomyces cerevisiae; E. coli strain: HB101. 32P-labeled probes recognized (G) GAPDH; (E) translation elongation factor EF-1α; and (O) outer membrane protein A sequences. (E and O were kindly provided by P. Philippsen, University of Basel, Switzerland and U. Henning, Max Planck Institute of Biology, Tübingen, Germany, respectively.) B. subtilis was not probed. M: 0.24–9.5 kb RNA ladder. 7.5 kb band (indicated) in embryo, Huh7, and HeLa cell lanes is a nuclear precursor RNA.
RNeasy Midi Spin column
RNeasy Midi spin column.
The RNeasy Midi spin column contained in the RNeasy Midi Kit.