RNeasy Micro Kit

細胞および組織サンプルからの最大45 μg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
  • 微量スタートサンプルからも一定した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 Micro Kitは少量の細胞や組織サンプルからRNA(最高45 µg)を精製するためにデザインされています。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。RNeasy Micro Kitを用いたRNA精製はQIAcube上で完全自動化が可能です。より多量のサンプルには、RNeasy Mini Kit(スピンカラム結合容量が100 µg RNA)、RNeasy Midi Kit(スピンカラム結合容量が1 mg RNA)、RNeasy Maxi Kit(スピンカラム結合容量が6 mg RNA)をご使用ください。
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RNeasy Micro Kit (50)
50 RNeasy MinElute Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free DNase I, Carrier RNA, RNase-free Reagents and Buffers
74004
$1,037.00
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The RNeasy Micro 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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Highly reproducible yields for sensitive applications.|High-quality total RNA from fine needle aspirates.|RNeasy Micro procedure.|Efficient on-column removal of genomic DNA.|Reliable RNA isolation from a single cell.|
Total RNA was isolated by 5 different users in duplicate from 2 mg of the indicated rat tissues, using the RNeasy Micro Kit. Quantitative, real-time RT-PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for c-jun.|A fine needle aspirate was obtained from rabbit liver using a 19.5-gauge biopsy needle, and the sample was immediately stabilized in an RNAprotect Tissue Tube. Total RNA was isolated using the RNeasy Micro Kit. The high quality of the RNA is shown by scanning with the Agilent 2100 Bioanalyzer.|The RNeasy Micro Kit is designed for isolation of total RNA from small samples. Optimized protocols are provided for RNA isolation from microdissected tissues, small amounts of standard tissues, fiber-rich tissues, and cells. Protocols are also available for RNA cleanup and concentration.|Total RNA was isolated from the indicated number of HeLa cells using the RNeasy Micro Kit. Quantitative, real-time PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe PCR Kit, with primers and probes specific for c-jun. In order to evaluate the efficiency of the on-column DNase digestion, each sample was amplified after reverse transcription (+RT) and directly, without a reverse transcription step (-RT). The lack of signal in the "-RT" samples indicates that no DNA was detectable. Of the "-RT" results, only the 50,000 cell data points are visible.|Total RNA was isolated from the indicated number of HeLa cells using the RNeasy Micro Kit. [A] Amplification plot. [B] Correlation of crossing point (CP) values and cell numbers. Real-time RT-PCR was carried out on the LightCycler System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for β-actin.|
Performance
RNeasy Micro Kitによって、少量のサンプルから極めて再現性の高いトータルRNA収量を実現できます。例えば単一細胞や少量の組織から、トータルRNAを確実に精製できます(図"単一細胞からの確実なRNA精製"、"高感度なアプリケーションでの高い再現性"、"穿刺吸引法サンプルからの高品質トータルRNA")。RNeasy Micro Kitは、カラム上でゲノムDNAを効率的に分解することによって、リアルタイムRT-PCRなどの定量的遺伝子発現解析で最高の感度を提供します(図"カラム上での効率的なゲノムDNA除去")。
Principle

RNeasy Micro Kitは、マイクロダイセクションで採取した組織および穿刺吸引物などの少量のサンプルからトータルRNAを精製するようにデザインされています。心臓および筋肉組織などのミリグラム単位の線維組織、および1個の細胞などの少数の細胞(例、FACSでの選別した細胞)からも精製可能です。マイクロダイセクションで採取したFFPE組織には、RNeasy FFPE Kitを推奨します。RNeasyテクノロジーでは、グアニジンイソチオシアネート溶解法と、シリカゲルメンブレンによる迅速な精製法を組み合わせることにより、トータルRNAの精製を簡便に行なうことが可能です。RNeasy MinElute Spin Columnのユニークなデザインによって、10 µlという最小限の溶出液量でRNAを濃縮できるようになっています。少ない溶出量で済むため、ダウンストリームアプリケーションにおいて反応液量を少量に保つことができ、反応効率が増加します。さらに、高純度なRNAは阻害物質を含まず、逆転写反応のようなダウンストリームアプリケーションに全溶出液を使用できます。

Procedure

RNeasy Micro Kitは少量のサンプルからトータルRNAを精製するためにデザインされています。まずサンプル(<5 mgの組織、または<5 x 105個の細胞)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 MinElute Spin Columnにアプライすると、RNA(最高45 µg)がシリカゲルメンブレンに結合します。RNeasy Micro Kitに含まれているRNase Free DNaseによって、高感度なアプリケーション向けのゲノムDNAをカラム上で簡単かつ効率的に分解することができます。DNaseと他の全ての夾雑物は効率的に洗い流され、最後に濃縮された高純度RNAが10~14 µlの水で溶出されます。

Applications

RNeasy Micro Kitを用いて少量の溶出量(10 µl)で精製したRNAは純度および再現性が高く、わずか1個の細胞を用いたリアルタイムRT-PCRのような高感度な遺伝子発現定量解析に最適です。

Feature
Specifications
Applications Northern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
Elution volume 10–14 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 <5 mg
Technology Silica technology
Time per run or per prep 30–40 minutes
Yield Varies

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Kit Handbooks
1
For purification of total RNA from small samples, including
  • animal and human cells
  • animal and human tissues
  • fibrous tissues
  • microdissected cryosections
and for RNA cleanup and concentration
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Images
Highly reproducible yields for sensitive applications.
Highly reproducible yields for sensitive applications.

Total RNA was isolated by 5 different users in duplicate from 2 mg of the indicated rat tissues, using the RNeasy Micro Kit. Quantitative, real-time RT-PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for c-jun.

High-quality total RNA from fine needle aspirates.
High-quality total RNA from fine needle aspirates.
A fine needle aspirate was obtained from rabbit liver using a 19.5-gauge biopsy needle, and the sample was immediately stabilized in an RNAprotect Tissue Tube. Total RNA was isolated using the RNeasy Micro Kit. The high quality of the RNA is shown by scanning with the Agilent 2100 Bioanalyzer.
RNeasy Micro Procedure
RNeasy Micro procedure.

The RNeasy Micro Kit is designed for isolation of total RNA from small samples. Optimized protocols are provided for RNA isolation from microdissected tissues, small amounts of standard tissues, fiber-rich tissues, and cells. Protocols are also available for RNA cleanup and concentration.

Efficient on-column removal of genomic DNA.
Efficient on-column removal of genomic DNA.
Total RNA was isolated from the indicated number of HeLa cells using the RNeasy Micro Kit. Quantitative, real-time PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe PCR Kit, with primers and probes specific for c-jun. In order to evaluate the efficiency of the on-column DNase digestion, each sample was amplified after reverse transcription (+RT) and directly, without a reverse transcription step (-RT). The lack of signal in the "-RT" samples indicates that no DNA was detectable. Of the "-RT" results, only the 50,000 cell data points are visible.
Reliable RNA isolation from a single cell.
Reliable RNA isolation from a single cell.
Total RNA was isolated from the indicated number of HeLa cells using the RNeasy Micro Kit. [A] Amplification plot. [B] Correlation of crossing point (CP) values and cell numbers. Real-time RT-PCR was carried out on the LightCycler System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for β-actin.