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

✓ オンライン注文による24時間年中無休の自動処理システム

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✓ 迅速で信頼性の高い（再）注文

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

￥94,000

研究用途専用。診断用ではありません。いかなる主張または表示も、疾病の診断、予防、治療のための情報を提供することを目的としてはいません。

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い（再）注文

特徴

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

製品詳細

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。

パフォーマンス

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 "")、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 ""、""、""）。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 ""）。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 ""）。

図参照

RNA yields from bone marrow samples.

RNA stability at room temperature.

RNA stabilized at 2–8°C.

High RNA integrity after freeze–thaw cycles.

Stability of 5 transcripts at room temperature.

Storage under different conditions.

原理

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。

操作手順

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。

アプリケーション

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

裏付けデータと数値

Stability of 5 transcripts at room temperature.

Specifications

Features	Specifications
Applications	RT-PCR, array analysis
Yield	11 µg
Time per run	95 min/12 samples
Main sample type	Bone marrow samples
Sample amount	2 ml
Processing	Manual (centrifugation)
Format	Spin column
Elution volume	80 µl (2 x 40 µl)
Technology	Silica technology

リソース

For collection, storage, and transport of bone marrow samples and stabilization and purification of intracellular RNA

Download Safety Data Sheets for QIAGEN product components.

Guenther et al., AMP 2006

Guenther et al., AACR 2007

This note is to inform you that the street address for PreAnalytiX GmbH has changed from “Feldbachstrasse” to “Garstligweg 8”. Please be informed that the update of the product labeling to the new address is ongoing.

In order to reduce paper consumption and oblige the growing number of customers requesting an environmentally friendly alternative to traditionally printed handbooks, we are now providing kit handbooks for Research Use Only (RUO) PreAnalytiX kits on our website only.

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

FAQ

You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .

FAQ ID - 3515

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

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

No. The elution buffer (Buffer BMR5) supplied in the PAXgene Bone Marrow RNA Kit must be used.

FAQ ID -2512

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

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

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

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

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

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

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

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

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

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

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

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

No. The PAXgene Bone Marrow RNA System has been optimized only for cellular RNA only.

FAQ ID -2507