AllPrep DNA/RNA/Protein Mini Kit

細胞および組織から DNA、RNA とタンパクを同時に分離精製

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AllPrep DNA/RNA/Protein Mini Kit (50)

Cat. No. / ID: 80004

For 50 preps: AllPrep DNA Spin Columns, RNeasy Spin Columns, Collection Tubes, RNase-Free Reagents and Buffers

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￥98,000

AllPrep DNA/RNA/Protein Mini Kitは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

貴重なサンプルから各分析対象物を最大量回収
簡単で効率的なプロトコールにより時間を節約
同一サンプルから分析物が得られ、データの比較が可能
フェノールやアセトンのような有機溶媒が不要

Product Details

AllPrep DNA/RNA/Protein Mini Kit は同一の細胞あるいは組織サンプルから同時にDNA、RNA、タンパク質を精製できます。本キットとAllprotect Tissue Reagentを組み合わせて使用することで、組織サンプルのDNA、RNA、タンパク質を迅速に安定化し、凍結することなく長期保存が可能になります。これらの組織はTissueRuptorあるいはTissueLyser IIを用いて効果的に破砕できます。

Performance

AllPrep DNA/RNA/Protein Mini Kitを用いて精製したゲノムDNAおよびトータルRNAは、リアルタイムPCRやリアルタイムRT-PCR解析に最適です（図 ""）。本キットを用いて精製したトータルタンパク質はSDS-PAGE およびウェスタンブロットに適しています（図）。

See figures

Reliable real-time PCR and RT-PCR analyses.

Reliable western blotting and SDS-PAGE.

Principle

AllPrep DNA/RNA/Protein Mini Kit によりシステムバイオロジーのためのサンプル調製を標準化できます。同一のサンプルからDNA、RNA、タンパク質を全て調製できるため、別々のサンプルからこれらの分析物を調製する際に起こるばらつきを排除できます。精製操作の前にサンプルを3つに分割する必要がなく、DNA、RNA、タンパク質を最も効率よく回収できます。さらに、フェノールやアセトンのような有機溶媒も不要です。

Procedure

最初にプロテアーゼ、DNase、RNaseを即座に不活性化するBuffer RLTで細胞あるいは組織を溶解し、ホモジナイズするので、分解のないDNA、RNA、タンパク質の分離ができます。ライセートをAllPrep DNA Spin cloumnにアプライし通過させます。高塩濃度バッファーと組み合わせることにより、カラムはゲノムDNAを選択的かつ効率的に結合させます。カラムを洗浄し、即使用できる高純度なDNAを溶出します。

AllPrep DNA Spin cloumnからのろ液にエタノールを添加することでRNAの最適な結合条件にし、このサンプルをRNeasy Spin cloumnにアプライします。トータルRNAはこのカラムのメンブレンに結合し、洗浄により夾雑物は効果的に除去されます。高品質なRNAをRNaseフリー水で溶出します。

RNeasy Spin cloumnからのろ液に、新規の水溶性タンパク質沈殿溶液であるBuffer APPを添加し、沈殿したタンパク質を遠心操作によりペレット化します。分解のないトータルタンパク質を適切なバッファーで再溶解した後、ダウンストリームアプリケーションにすぐに使用できます。本キットにはタンパク質ペレットを溶解しSDS-PAGEに使用できるBuffer ALOが入っています（フローチャート " AllPrep DNA/RNA/Protein 操作手順"）。

See figures

AllPrep DNA/RNA/Protein procedure.

Applications

AllPrep DNA/RNA/Protein Mini Kit は、同一の細胞や組織サンプルからDNA、トータルRNA、タンパク質を同時に精製し、効率的なサンプル調製を実現します。本キットを用いて精製したトータルタンパク質はSDS-PAGE およびウェスタンブロットに最適です。AllPrep DNA/RNA/Protein Mini Kit を用いて精製したゲノムDNAおよびトータルRNAは以下のようなアプリケーションを含む全てのアプリケーションに最適です。

リアルタイムPCR解析
リアルタイムRT-PCR解析

Supporting data and figures

Reliable real-time PCR and RT-PCR analyses.

Rat tissues were stabilized in Allprotect Reagent, and DNA and RNA were purified using the AllPrep DNA/RNA/Protein Mini Kit. The C_T values in real-time PCR and real-time RT-PCR analyses were similar to those achieved with tissues stabilized in liquid nitrogen.

Specifications

Features	Specifications
Applications	PCR, RT-PCR,real-time PCR, real-time RT-PCR, Southern-,Northern-,Western Blotting, Microarrays, Genotyping,1D and 2D gel electrophoresis
Main sample type	Easy-to-lyse tissue or cells
Elution volume	RNA: 30 - 50µl, DNA: 100 - 200µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Total RNA, genomic DNA and western-grade proteins
Format	Spin column
Yield	varies, depends on sample type
Time per run or per prep	60 min
Processing	Manual (centrifugation)
Technology	Silica technology, Protein precipitation
Sample amount	up to 30 mg tissue or 1 x 10e7 cells

Resources

For simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample

For simultaneous purification of genomic DNA, total RNA and total protein from the same cell or tissue sample

Download Safety Data Sheets for QIAGEN product components.

For simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample

For simultaneous purification of genomic DNA, total RNA and total protein from the same cell or tissue sample

FAQ

The AllPrep DNA/RNA/Protein Mini Kit is optimized for use with easy-to-lyse tissues. For fibrous (e.g., heart) and lipid tissues (e.g., brain), we recommend using a portion of the tissue sample for RNA purification with the RNeasy Fibrous Tissue Mini Kit and RNeasy Lipid Tissue Mini Kit, respectively.

FAQ ID -1579

The detailed composition of Buffer ALO is confidential. Buffer ALO is part of the AllPrep DNA/RNA/ Protein Kit and functions as protein resuspension buffer. Buffer ALO is a Laemmli-related sample buffer for use in SDS-PAGE. In addition to other reagents, Buffer ALO comprises SDS, bromophenol blue, and dithiothreitol (DTT). DTT is added by the user prior the first use of the kit.

FAQ ID -2805

No, Buffer APP in the AllPrep DNA/RNA/Protein Mini Kit is a novel aqueous protein precipitation solution and has some advantages over acetone with regard to recovery of small proteins (about 10 kDa and smaller).

FAQ ID -1576

Yield and integrity of genomic DNA depend on the disruption and homogenization method used with the Allprep DNA/RNA Kits.

Homogenization with the TissueRuptor (or other rotor–stator homogenizer, such as the Polytron) or the TissueLyser results in greater DNA fragmentation (depending on homogenization time and intensity). However, shorter DNA fragments are easier to elute, so DNA yields will be higher.

In contast, homogenization using the QIAshredder (or a syringe and needle) is more gentle, resulting in less DNA fragmentation. However, longer DNA fragments are more difficult to elute, so DNA yields may be lower.

FAQ ID -1751

In the protein precipitation procedure of the AllPrep DNA/RNA/Protein protocol, be sure to dissolve the protein pellet completely in Buffer ALO or 1x SDS-PAGE sample buffer.

For easier dissolving of proteins, dissolve the pellet in 5% (w/v) SDS or 8 M urea. Commonly, there is insoluble material left even after vigorous shaking or heating. This is typical for all kinds of protein precipitation under heavily denaturing conditions.

Briefly centrifuge the sample and use the supernatant for downstream analysis.

FAQ ID -1580

The exact composition of Buffer APP is confidential. Buffer APP is a component of the AllPrep DNA/RNA Protein Kit and functions as a protein precipitation buffer, containing, among other components, zinc chloride.

FAQ ID -2804

Yes, please follow the Supplementary Protocol Acetone precipitation of protein from Buffer RLT or Buffer RLT Plus lysates (RY22).

Important Note:

Do not use TCA to precipitate protein from Buffer RLT and Buffer RLT Plus lysates. These buffers contain guanidine thiocyanate, which can form highly reactive compounds when combined with acidic solutions.

For simultaneous purification of DNA, RNA, and protein from the same sample (either cultured cells or easy-to-lyse tissues), we recommend using the AllPrep DNA/RNA/Protein Mini Kit. This kit allows precipitation of protein from Buffer RLT lysates using a novel protein precipitation buffer, Buffer APP.

Please note that Buffer APP is not compatible with Buffer RLT Plus. Acetone should be used instead to precipitate protein from RLT Plus lysates.

FAQ ID -1164

No, Buffer APP for protein precipitation in the Allprep DNA/RNA/protein Mini Kit is not compatible with Buffer RLT Plus. Acetone can be used to precipitate proteins from samples homogenized in Buffer RLT Plus contained in the Allprep DNA/RNA Mini Kit and RNeasy Plus Mini Kit.

FAQ ID -1577

Buffer RLT Plus contains detergents that are more ideal for binding of DNA to the AllPrep DNA mini spin columns.

But RLT Plus is not compatible with Buffer APP used to precipitate proteins in AllPrep DNA/RNA/Protein Mini kit. Therefore, we chose RLT instead of RLT Plus to get both DNA and proteins with this kit.

RLT lysates can also be used in AllPrep RNA/DNA kit or the AllPrep DNA/RNA Micro kit. DNA and RNA yields should be fine, but RNA may contain slightly higher levels of genomic DNA contamination.

FAQ ID - 3391

A custom protocol can be requested through the online QIAcube protocol request page: <https://www.qiagen.com/qiacube/customized/protocoladd.aspx >.

FAQ ID - 3323