RNeasy Protect Animal Blood System

動物血液の採取と安定化およびRNA/miRNA の精製

S_1084_8_GEN_Kitrot

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RNAprotect Animal Blood Tubes (50 x 100 µl)

Cat. No. / ID:  76544

50 tubes for collection, storage, and transport of 100 µl animal blood samples with RNA stabilization; to be used with the RNeasy Protect Animal Blood Kit
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¥23,000
TubeKit
RNAprotect Animal Blood Tube
RNeasy Protect Animal Blood Kit
Quantity
50 x 100 µl
50 x 500 µl
RNeasy Protect Animal Blood System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

  • 動物血液の信頼できる遺伝子発現解析
  • 少量血液の簡便な採取や保存
  • 高品質トータルRNAを迅速に精製
  • miRNAの効率的な回収

Product Details

RNeasy Protect Animal Blood System は、動物血液から高品質なRNAを調製するためのトータルソリューションを実現します。RNAprotect Animal Blood Tube に採取したマウス、ラット、その他小動物からの血液中の細胞性RNAは、チューブ内の試薬により迅速に安定化されます。続いてRNeasy Protect Animal Blood Kit を用いてトータルRNAを精製します。RNeasy Protect Animal Blood Kit とRNAprotect Animal Blood Tubes は個別にご注文ください。

また、microRNA(miRNA)を精製する際は、RNeasy Protect Animal Blood Kit を使用することにより、低分子RNAを含むトータルRNA画分や低分子RNAのみの濃縮画分を得ることができます。miRNAを含むトータルRNAの精製には、Buffer RWT(Cat.no. 1067933)も必要です。miRNA 濃縮画分とトータルRNA(200 nt以上)画分を別々に分離精製する場合には、RNeasy MinElute Cleanup Kit(Cat.no. 74204)が必要です。

Performance

RNAprotect Animal Blood Tube に採取した血液サンプルは、常温での輸送や冷蔵での保存を簡単に行なえます。サンプルは15 ~ 25 ℃で最高48 時間、2 ~ 8 ℃で最高2 週間、-20℃で少なくとも3 ヶ月保存可能で、有意なRNA分解は観察されません(図 " インタクトなRNA")。RNAprotect Animal Blood Tube に採取した動物血液では転写レベルでの変化が抑えられ、この結果、リアルタイムRT-PCRやマイクロアレイ解析のようなアプリケーションで信頼できる遺伝子発現解析を実現します(図 " 転写物を効率的に安定化")。RNeasy Protect Animal Blood Kit によりRNA収量は再現性が高く、30 分未満のマニュアルでの作業で調製は完了します(図 " 再現性のあるRNA収量")。実績のあるRNeasy テクノロジーにより、ゲノムDNAの混入がない、高純度なRNAが得られます。
See figures

Principle

血液サンプル採取時、遺伝子発現プロファイルは採血後数分で大幅に変化することがあります。EDTAなどの抗凝固剤は血液の凝固を防ぎますが、転写物の誘導やmRNAターンオーバーの抑制により生じる遺伝子発現の変化は避けられません。RNAprotect Animal Blood Tubesで動物血液を採取することにより、転写物レベルの変化を防ぐことができます。安定化した血液を処理・ホモジナイズした後、実績あるRNeasyシリカメンブレンテクノロジーでサンプルからRNAを精製します(図 " RNeasy Protect Animal Blood フローチャート")。
See figures

Procedure

血液サンプルをRNAprotect Animal Blood Tubes に採取します。チューブ内には、赤血球を溶解し細胞内RNAを安定化する試薬が入っています。RNAprotect Animal Blood Tubesを遠心してサンプルをペレット化し、水で洗浄してBuffer RSBに再懸濁します。Buffer RBT中でproteinase K処理した後、サンプルをQIAshredder spin columnsに通し、遠心によりホモジナイズします。サンプルにエタノールを加えた後、サンプルをRNeasy MinElute spin columnsに通し、遠心します。結合したトータルDNAをDNase処理し、Buffer RW1およびBuffer RPEで順に洗浄します。Buffer REBで溶出することにより、純度の高いRNAが得られます。RNeasy Protect Animal Blood Kitを用いたRNA精製はQIAcube上で完全に自動化できます。miRNAを含むトータルRNAの精製用に、洗浄用のBuffer RWT(別売り、カタログ番号 1067933)を用いた別プロトコールが用意されています。低分子RNA(miRNAを含む)と高分子RNA(200 nt以上)を2つの分画に分けて精製する場合には、RNeasy MinElute Cleanup Kitも必要です。

Applications

RNeasy Protect Animal Blood Systemで精製したトータルRNAおよびmiRNAは次のようなアプリケーションに最適です。

遺伝子発現解析
RT-PCR
リアルタイム定量RT-PCR
ディファレンシャルディスプレイ
cDNA合成

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsNorthern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
Elution volume14 µl
Sample amount100 µl or 500 µl animal blood
Main sample typeTissue, cells
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinTotal RNA
ProcessingManual (centrifugation)
FormatSpin column
TechnologySilica technology
YieldRat blood (500 μl): 10–30 µg; Mouse blood (100 μl): 4–8 µg

FAQ

Do you have a kit for RNA isolation from any kind of sample type?

The RNeasy 96 Universal Tissue Kit enables high-throughput purification of RNA from any animal or human tissue sample, including difficult-to-lyse fibrous and fatty tissues. For single tube format RNA purification, the RNeasy Plus Universal Kit is also available. 

Please refer to the Selection guide for RNA isolation for all sample types to find the optimal solution for your sample source.

FAQ ID -627
What is the composition of Buffer RLT?

The exact composition of Buffer RLT is confidential. This buffer is a proprietary component of RNeasy Kits. Buffer RLT contains a high concentration of guanidine isothiocycanate, which supports the binding of RNA to the silica membrane. Buffer RLT can be purchased separately (cat. no. 79216)

Note: note that ß-mercaptoethanol should be added to Buffer RLT before use to effectively inactivate RNAses in the lysate (10 µl ß-Mercaptoethanol per 1 ml Buffer RLT).

FAQ ID -2793
How can I check the integrity of RNA purified using RNeasy Kits?

The integrity and size distribution of total RNA purified with RNeasy Kits can be checked by denaturing-agarose gel electrophoresis, the Agilent 2100 bioanalyzer, or the QIAxcel Advanced System with the QIAxcel RNA QC Kit v2.0.

The respective ribosomal species should appear as sharp bands on the stained gel. 28S ribosomal RNA bands should be present with an intensity approximately twice that of the 18S RNA band. If the ribosomal bands are not sharp, but appear as a smear of smaller sized RNAs, it is likely that the RNA sample has suffered major degradation during preparation.

Size of ribosomal RNAs from various sources

Source

rRNA

Size (kb)

E. coli

16S

1.5

 

23S

2.9

S. cerevisiae

18S

2.0

 

26S

3.8

Mouse

18S

1.9

 

28S

4.7

Human

18S

1.9

 

28S

5.0

 

 

 

 

 

 

 

 

 

FAQ ID -1024
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
What are the effects of low A260/A230 ratios in RNA preparations on downstream applications?

The efficiency of downstream applications depends strongly on the purity of the RNA sample used.  Pure RNA should yield an A260/A230 ratio of around 2 or slightly above; however, there is no consensus on the acceptable lower limit of this ratio.  Possible candidates that can increase the A230 include “salt”, carbohydrates, peptides, and phenol (or aromatic compounds in general).  In our experience, the increased absorbance at 230 nm in RNA samples is almost always due to contamination with guanidine thiocyanate, present at very high concentrations in the lysis buffer or extraction reagent used in most RNA purification procedures.

Please find an article discussing the effect of low 260/230 ratios in RNA preparations on downstream applications on page 7 of QIAGEN Newsletter March 15, 2010 . In summary, we found that concentrations of guanidine thiocyanate of up to 100 mM in an RNA sample do not compromise the reliability of downstream applications.

 

 

FAQ ID -2248
What is the composition of Buffer RPE?
The exact composition of Buffer RPE is confidential. Buffer RPE is a mild washing buffer, and a proprietary component of RNeasy Kits. Its main function is to remove traces of salts, which are still on the column due to buffers used earlier in the protocol. Ethanol, which is added by the user just before using the kit for the first time, is an important ingredient of Buffer RPE.
FAQ ID -2797
What is the composition of Buffer RW1?

The exact composition of Buffer RW1 is confidential. Buffer RW1 is a proprietary component of RNeasy Kits. Buffer RW1 contains a guanidine salt, as well as ethanol, and is used as a stringent washing buffer that efficiently removes biomolecules such as carbohydrates, proteins, fatty acids etc., that are non-specifically bound to the silica membrane. At the same time, RNA molecules larger than 200 bases remain bound to the column.

Note: Buffer RW1 should not be used for isolation of small RNAs, for example, microRNAs or fragmented RNA from formalin-fixed tissues, as these smaller fragments will be washed away.  Buffer RWT should be used instead.

FAQ ID -2796