PAXgene Bone Marrow RNA Kit

骨髄液検体の保存と輸送および細胞内RNAの安定化と精製

S_1392_RPA_PAX1166

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PAXgene Bone Marrow RNA Kit (30)

Cat. No. / ID:  764133

For 30 RNA preps: 30 PAXgene Spin Columns, 30 PAXgene Shredder Spin Columns, Processing Tubes, RNase-free DNase I, RNase-free reagents and buffers. To be used in conjunction with PAXgene Bone Marrow RNA Tubes
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$879.00
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✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 保存、安定化、精製を統合したシステム
  • 検体採取後即座に細胞内RNAを安定化
  • 様々な骨髄液検体から迅速な精製
  • 簡便かつ安全な輸送

Product Details

PAXgene Bone Marrow RNA System は骨髄液の安定化、保存、輸送のためのPAXgene Bone Marrow RNA Tube と、シリカメンブレンを利用したスピンカラムフォーマットのRNA分離精製用PAXgene Bone Marrow RNA Kit で構成されています。汎用されている抗凝固剤を用いて一般的な方法で骨髄を採取します。2 ml の検体を即座にPAXgene Bone Marrow RNA Tube に移すことで検体中のRNAは安定化され、室温、冷蔵あるいは凍結して保存できます。後日PAXgene Bone Marrow RNA Kit を用いてRNAを精製することも、–20℃あるいは–70℃で保存することも可能です。

Performance

PAXgene Bone Marrow RNA Kitは、多様なヒト骨髄液検体から細胞RNAを効率的に分離・精製し (図 " 骨髄液検体からのRNA収量")、細胞を多く含む粘性の高い検体にも至適化されています。後日RNAを精製することが可能で、室温、冷蔵、冷凍保存したサンプルからPAXgene Bone Marrow RNA Kitを用いて劣化のない質の高いRNAを精製できます(図 " 室温でのRNA安定性"、" 2~8℃で保存中のRNA安定性"、" 凍結融解後も分解していないRNA")。RNA Tubesは、RNA分子がRNasesにより分解するのを防ぎ、生体外での変化を最小に抑えるため正確な遺伝子発現解析が行なえます(図 " 室温での5種類の転写物の安定性")。また、DNase処理が含まれているので、ゲノムDNAの混入は最小限に抑えられます。弊社および外部施設での検討で、分離・精製した検体(n=157)の95%でゲノムDNAの残存率が0.85%未満という結果が得られました。A260/A280 値は1.8〜2.2であり、RNAが高純度であることを示しています(図 " 種々の条件下で保存")。
See figures

Principle

骨髄液中の個々のmRNA種のコピー数は、室温での保存や輸送中に著しく変動するため、信頼できる遺伝子発現研究は非常に困難になっています。PAXgene Bone Marrow RNA Tubes は、RNA 分子が分解しないよう保護し、サンプル中のRNAを即座に安定化するため、サンプルを室温、冷蔵、冷凍で保存できます。

Procedure

汎用されている抗凝固剤を用いて一般的な方法で骨髄を採取します。次に、2 ml のサンプルをPAXgene Bone Marrow RNA Tubeに移します。サンプル中のRNAは即座に安定化され、室温、冷蔵、冷凍で保存できるようになります。PAXgene Bone Marrow RNA Kitを用いると、ヒト骨髄液検体から細胞RNAの分離/精製を迅速かつ容易に行なうことができます。本キットはスピンカラムフォーマットでシリカメンブレンを使用しており、インタクトな高品質RNAを効率的に精製することが可能です。

Applications

PAXgene Bone Marrow RNA Kit を用いて精製したRNAは次のような様々なダウンストリームアプリケーションに直ちに使用できます。

  • 特殊な転写物のリアルタイムRT-PCR解析
  • 複雑な発現プロファイルのアレイ解析

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsRT-PCR, array analysis
Yield11 µg
Time per run95 min/12 samples
Main sample typeBone marrow samples
Sample amount2 ml
ProcessingManual (centrifugation)
FormatSpin column
Elution volume80 µl (2 x 40 µl)
TechnologySilica technology

Resources

パンフレット (3)
For collection, storage, and transport of bone marrow samples and stabilization and purification of intracellular RNA

 

MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
キットハンドブック (2)
For the collection of human bone marrow and immediate stabilization of intracellular RNA
For isolation and purification of intracellular RNA from whole bone
marrow samples stabilized in PAXgene Bone Marrow RNA Tubes
Safety Data Sheets (1)
Certificates of Analysis (1)
Brochures & Guides (2)
For collection, storage, and transport of bone marrow samples and stabilization and purification of intracellular RNA

 

Gene Expression Analysis (1)
Kit Handbooks (2)
For isolation and purification of intracellular RNA from whole bone
marrow samples stabilized in PAXgene Bone Marrow RNA Tubes
For the collection of human bone marrow and immediate stabilization of intracellular RNA

FAQ

Where can I find additional information for PreAnalytiX PAXgene products?
You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .
FAQ ID - 3515
How much time may the aspirated bone marrow sample be allowed to incubate before pipetting it into the PAXgene Bone Marrow RNA Tube?
As gene expression starts to change in a sample removed from the human body within minutes, we highly recommend transferring the samples within a few minutes into the stabilization solution of the PAXgene Bone Marrow RNA tubes for cell lysis directly linked to RNA stabilization to take place.
FAQ ID -2504
I routinely heat an aliquot of the RNA eluate prior to my RT reaction. Do I still need to heat the eluate after the PAXgene Bone Marrow RNA preparation?
Yes. Heating temperature and incubation time will vary depending on different RT protocols. Therefore, the final heating step should be performed routinely at the end of the protocol.
FAQ ID -2513
Can I use water to elute the RNA in the PAXgene Bone Marrow RNA Kit?
No. The elution buffer (Buffer BMR5) supplied in the PAXgene Bone Marrow RNA Kit must be used.
FAQ ID -2512
Can specific types of white blood cells be enriched from a PAXgene Bone Marrow RNA Tube prior to the RNA isolation?
No. The cells are lysed in the PAXgene Bone Marrow RNA Tube. There is no method available to enrich for certain sub-populations of cells.
FAQ ID -2508
How are the supernatants resulting from the first two spin steps of the PAXgene Bone Marrow RNA kit purification procedure disposed?

The supernatants resulting from the first two spin steps of the PAXgene Bone Marrow RNA Kit purification procedure may still contain biologically hazardous material. We recommend classifying the tube-processed supernatant of the kit as both a biological and chemical waste. Supernatants can be decontaminated by autoclaving and disposed of as chemical waste.  Disposal should be done in accordance with local, state and federal regulations.

CAUTION: Do not add bleach or acidic solutions directly to the sample preparation waste.

FAQ ID -2516
Can the PAXgene Bone Marrow RNA System be used with animal bone marrow or with blood samples?
The PAXgene Bone Marrow RNA System was developed for intracellular RNA isolation from human bone marrow samples. Bone marrow samples from other species or from blood were not tested.
FAQ ID -2501
Can bone marrow from leukemia patients with elevated numbers of white blood cells be processed with the PAXgene Bone Marrow RNA System?
Yes. We have tested the PAXgene Bone Marrow RNA System with bone marrow samples containing very high numbers of leukocytes. Bone marrow samples with higher leukocyte numbers are indicated by a highly viscous solution after addition of buffers BMR1 and BMR2 followed by incubation. These challenging samples are further diluted according to descriptions given in the protocol and do not lead to problems in the extraction procedure.
FAQ ID -2510
Can the PAXgene RNA stabilization solution/bone marrow mixture from the PAXgene Bone Marrow RNA Tube be disposed of by adding bleach?
Yes. You may dispose of the waste product from the PAXgene Bone Marrow RNA Tube with a 5% sodium hypochlorite bleach solution, in a 1:9 ratio (1 part bleach : 9 parts tube contents).
FAQ ID -2515
How should the PAXgene Bone Marrow RNA Tubes be frozen after collection and for how long?

The PAXgene Bone Marrow RNA Tubes should be frozen at –20°C and –70°C in a wire rack, not in an expanded polystyrene foam tray.

If the PAXgene Bone Marrow RNA Tubes are to be stored at temperatures lower than –20ºC, freeze them first for 24 h at –20ºC, then transfer them to –70ºC.

Currently available data shows stabilization of cellular RNA for several months at –20°C or –70°C. For more information from ongoing studies evaluating stability for longer time periods, please contact QIAGEN Technical Services.

 

 

FAQ ID -2505
What RNA yields are expected with the PAXgene Bone Marrow RNA System?

Bone marrow samples are extremely heterogeneous, consisting of varying amounts of cells, tissue, and solid matter. RNA yields are highly donor-dependent and can vary greatly from sample to sample as well as between replicates from the same donor due to sample-intrinsic heterogeneity.

In a study of 295 bone marrow samples, yields of RNA isolated from 2 mL human bone marrow were between 2.0 and 44.7 µg (5-95% quantiles, mean = 15.8 µg, median = 11.0 µg).

FAQ ID -2506
Is RNA from red blood cells also isolated when RNA is extracted from a PAXgene Bone Marrow RNA Tube?
Yes. The PAXgene Bone Marrow RNA System purifies RNA from whole bone marrow. Therefore it will also purify RNA from other cells, e.g. reticulocytes and RBCs. Array analyses have shown higher globin mRNA levels for RNA isolated from whole bone marrow compared to RNA isolated from mononuclear cells.
FAQ ID -2509
Is the 260/230 nm absorbance ratio specified for RNA derived from the PAXgene Bone Marrow RNA System?

No. The 260/230 nm absorbance ratio is not specified for the PAXgene Bone Marrow RNA extraction kit. In contrast to a reduced 260/280 ratio, which results from a protein contamination of the eluted RNA and can lead to inhibitory effects in down stream applications, there is no negative impact of a reduced 260/230 ratio on any down stream application reported to date and therefore it is not necessary to specify this.

In addition, because the elution buffer contains very low concentrations of salt that absorb at wavelengths between 220 and 230 nm, it is necessary to zero the photometer properly. Do this by preparing a solution for zeroing the photometer which contains a similar salt concentration as the sample to be measured by using the same volumes. Add the same volume of Buffer BMR5 as the volume of RNA sample to be diluted into a fresh volume of the buffer in which you will measure the absorption (we recommend 10 mM Tris Cl pH 7.5 buffer for this purpose).

FAQ ID -2514
What is in the elution buffer in the PAXgene Bone Marrow RNA kit?
The PAXgene Bone Marrow RNA kit elution buffer (Buffer BMR5) is a proprietary low-salt buffer. In combination with the final heat denaturation step, it leads to optimal performance of the isolated RNA in RT-reactions. There is no interference with downstream applications reported to date.
FAQ ID -2511
Can the PAXgene Bone Marrow RNA System be used for other human body fluids than bone marrow samples or cultured cells of human origin?
No. The PAXgene Bone Marrow RNA System was developed for intracellular RNA isolation from human bone marrow samples. PreAnalytiX offers a dedicated collection and RNA extraction system for human blood (see the PAXgene Blood RNA System). QIAGEN also offers RNA stabilization and isolation systems for saliva and cultured cells. Please contact QIAGEN technical service for more information or visit www.qiagen.com.
FAQ ID -2502
Can bone marrow aliquots other than 2 mL be pipetted into the PAXgene Bone Marrow RNA Tube?
No. Transferring bone marrow volumes different than 2mL into the PAXgene Bone Marrow RNA Tube is not recommended. The tube and its contents were optimized for performance with 2 mL of bone marrow. The use of other volumes is not recommended.
FAQ ID -2503
Can the PAXgene Bone Marrow RNA System be used to isolate genomic DNA or viral RNA and DNA?
No. The PAXgene Bone Marrow RNA System has been optimized only for cellular RNA only.
FAQ ID -2507