TissueLyser II

48あるいは192サンプルをビーズで破砕

Features

簡便で安全な破砕方法
ハイスループット破砕に最適なアダプターセット
様々な付属品が入手可能
溶解困難な組織から再現性の高い結果を実現
QIAGEN 自動化用製品に対応

Product Details

TissueLyser II はステンレススチール、タングステンカーバイトあるいはガラスビーズと共に生体サンプルをプラスチックチューブ中で高速で振蘯することにより複数の生体サンプルの同時破砕を実現します。適切なアダプターセットを用いて48あるいは192サンプルまで同時に処理できます。また、大量サンプルの処理には破砕ジャーセットを使用できます。様々なビーズ、ビーズ・ディスペンサー、コレクション・マイクロチューブやキャップも入手できます。アダプターセットを含むTissueLyser IIの付属品はTissueLyser（販売終了）でも使用可能です。

Performance

TissueLyser II はヒト、動物、あるいは植物の組織、バクテリア、酵母などのハイスループット破砕に最適です。溶解困難な組織からでさえも、再現性のある高品質なDNA、RNA、miRNA、タンパク質精製が可能です（図 “動物組織の効果的な破砕”、“植物組織からの再現性の高いRNA精製”、“”、“”、“”、および“”）。

See figures

Intact protein suitable for all types of analysis.

High-quality DNA from animal tissues.

High-quality DNA from plant tissues.

Reliable detection of miRNAs.

Principle

ジェノタイピング、遺伝子発現解析、プロテオミクスにおける実験ではDNA、RNA、タンパク質を確実に高収量で得るために生体サンプルを効果的に破砕する必要があります。TissueLyser IIシステムを用いたサンプルとビーズの高速なシェーキングにより、サンプルがビーズにより打ち砕かれ、すり潰されて、容易に破砕されます。TissueLyser システムは生体分子を完全に遊離するため、迅速かつ完璧にサンプルを破砕し、さらにQIAGEN 製品 (表 “QIAGEN 破砕システムに対応するQIAGEN 精製用キット”)を用いて容易に精製を行なえるよう、同時にサンプルをホモジナイズします。

QIAGEN はサンプル採取からDNA、RNA、タンパク質の精製および解析までのトータルソリューションを提供しており、TissueLyser IIはその破砕工程を構成しています。TissueLyser II はQIAsymphony SP のようなQIAGEN のハイスループットなサンプル調製および解析の自動化プロセスの前処理に最適です (表 “QIAGEN によるハイスループットな自動化”)。この使い易い装置は1 ～ 96 サンプルからのDNA、RNA、タンパク質精製を自動化します。TissueLyser II はマニュアルのQIAGEN サンプル調製用キットにも使用可能です (表 “QIAGEN 破砕システムに対応するQIAGEN 精製用キット”)。

RNA アプリケーションでは、新鮮な組織を RNAlater RNA Stabilization Reagent 中で安定化させることにより、サンプル取り扱い中のRNA 分解を回避できます。DNA、RNA、タンパク質の精製を必要とするアプリケーションでは、これら3 種類を迅速に安定化するために Allprotect Tissue Reagent で新鮮な組織を処理します。

QIAGENによるハイスループットな自動化
装置	目的	スループット
QIAsymphony SP	DNA、RNA、タンパク質の精製	1～96サンプル／ラン
BioRobot Universal System	DNA、RNAの精製	96ウエルフォーマット
QIAxcel	DNAフラグメントおよびRNAの電気泳動解析	最大96サンプル／ラン
QIAgility	反応セットアップ	最大384サンプル／ラン
Rotor-Gene Q	リアルタイムPCRとHRM（high-resolution melting）解析	最大100サンプル／ラン
PyroMark Q96 ID	メチル化および突然変異解析	最大96サンプル／ラン

QIAGEN破砕システムに対応するQIAGEN精製用キット
精製する生体物質	サンプルタイプ	QIAGEN キット
RNA	溶解が容易な組織	RNeasy Kits
		RNeasy Plus Kits
		RNeasy Protect Kits
RNA	繊維性組織	RNeasy Fibrous Tissue Kits
RNA	全タイプの組織	RNeasy Lipid Tissue Kits
RNA	全タイプの組織	RNeasy Universal Tissue Kits
RNA	植物組織	RNeasy Plant Mini Kit
RNA	酵母	RNeasy Kits
RNA	バクテリア	RNeasy Protect Bacteria Kits
microRNA	全タイプの組織	miRNeasy Kits
DNA	ヒト組織	QIAamp DNA Kits
DNA	動物組織	DNeasy Blood & Tissue Kits
タンパク質	組織	Qproteome Mammalian Protein Prep Kit
リン酸化タンパク質	組織	PhosphoProtein Purification Kit
糖タンパク質	組織	Qproteome Glycoprotein Kits
DNA およびRNA	組織	AllPrep DNA/RNA Kits
DNA, RNA, タンパク質	組織	AllPrep DNA/RNA/Protein Mini Kit

Procedure

TissueLyser Adapter Sets により、コレクションマイクロチューブ中で最大2 x 96個のサンプルを、2 mlマイクロ遠心チューブで最大2 x 24サンプルを処理します。溶解バッファーなしにサンプルを破砕する場合は、前もってアダプターを-80 ℃で冷却できます。破砕中にチューブを密封することによりクロスコンタミを回避することは、リアルタイムRT-PCRやマイクロアレイ解析のような高感度なダウンストリームアプリケーションでは特に重要です。サンプルのタイプにより、ステンレス製、タングステンカーバイド製、ガラス製のビーズで破砕をします。マイクロ遠心チューブにビーズ1個を入れるため、あるいはコレクションマイクロチューブに96個のビーズを並行して入れるための TissueLyser Bead Dispensers もお届けしています。TissueLyser IIはまた Grinding Jar Setsと組み合わせて、大量サンプルの破砕も可能です。

Applications

ランあたり最大192サンプル処理が可能なTissueLyser IIは、ゲノミクス、トランスクリプトミクス、プロテオミクスのアプリケーションに必要な生体情報を獲得する際に、初期の工程で理想的なツールです。次世代ハイスループットシークエンシングテクノロジーにとっては、 TissueLyser IIは最適な破砕装置です。TissueLyser IIは様々なQIAGENサンプル精製キットと互換性があり、サンプル精製はマニュアル、あるいはQIAcube、QIAsymphony SP、QIAxtractor、EZ1 Advanced、EZ1 Advanced XLでの自動化により行なえます。

Supporting data and figures

Reproducible RNA purification from plant tissues.

Frozen plant leaves were disrupted using the TissueLyser II (2 x 1 minute). RNA was purified using the RNeasy Plant Mini Kit and analyzed on a 1.2 % formaldehyde agarose gel. The ribosomal RNA bands were sharp and of equal intensity, indicating reproducible purification of intact RNA. T: tomato (100 mg); A: arabidopsis (25 mg); C: cotton (100 mg); M: maize (100 mg); R: rape (100 mg).

Specifications

Features	Specifications
Disruption principle
Typical run time
Technical data
Kits compatible with instrument
Protocol/main application on this intrument
Throughput
Features
Technology

Resources

FAQ

Efficient DNA isolation requires thorough sample disruption and digestion.

Although the QIAamp and DNeasy procedures requires no mechanical disruption of the tissue sample, the lysis time will be reduced if the sample is ground in liquid nitrogen or mechanically homogenized in advance. For mechanical homogenization, a rotor–stator homogenizer, such as the QIAGEN TissueRuptor, or a bead mill, such as the QIAGEN TissueLyser, can be used.

To improve digestion of tough tissue samples, Proteinase K incubation at 56°C can be performed overnight. DNA yields may be improved by increasing the amount of Proteinase K or by adding additional proteinase K after several hours of digestion.

FAQ ID -374

Buffer ATL can produce foam when used in conjunction with the TissueRuptor. You can add 0.5% final volume of Reagent DX (cat. no. 19088) to prevent the formation of foam.

FAQ ID -9143

All TissueLyser II repairs must be done by QIAGEN’s Regional Repair Center. Please contact QIAGEN Technical Service.

FAQ ID -9169

Frequency can be set from 3–30 Hz and is kept constant by a speed controller during operation. It may vary in a range of 0.2–0.4 Hz. If you see larger variations, please make sure not to overload the sample tubes with too much starting material.

If the issue persists, please contact QIAGEN Technical Service to have a repair by QIAGEN’s Regional Repair Center.

FAQ ID -9170

Check assembly of the TissueLyser II adapter. It is very important to assemble the TissueLyser adapter correctly. If done incorrectly, the caps of tubes will not remain tight during the disruption and can result in the leak.

FAQ ID -9171

When a new TissueLyser II is shipped, it has a transport security device located on the underside of the TissueLyser II. This is comprised of 2 screws as shown below. Remove the 2 screws using a screwdriver. Do not discard the screws, as they will need to be reinstalled if the TissueLyser II needs to be transported in the future.

FAQ ID -9172

We do not recommend using the adapters of the TissueLyser II with liquid nitrogen. Instead, the adapters can be precooled at –80°C, either in the freezer or on dry ice.

FAQ ID -9173

We recommend using the stainless steel Grinding Jar Set (cat. no. 69985) for disruption of tough samples, such as bone and teeth. Best results were seen with:
1. Take no more than 1 g of bone
2. Add material to the Grinding Jar Set containing a grinding ball
3. Pour in liquid nitrogen
4. As soon as the liquid nitrogen evaporates, assemble the Grinding Jar Set into the TissueLyser
5. Operate for 1 min at 30 Hz

FAQ ID -9174

We have not specifically tested this application. We suggest using a 7 mm stainless steel bead and disrupting the samples twice at 30 Hz for 2 min each.

Please be advised that these are suggestions only and have not been thoroughly tested by QIAGEN. It is the user’s responsibility to test and further optimize if necessary.

FAQ ID -9175

We do not have instructions or recommendations for you to perform frequency verification on our TissueLyser II. However, our Regional Repair Center can offer this as a service. Please contact QIAGEN Technical Service to request service.

FAQ ID -9176

We have not specifically tested this application. However, several customers have successfully used QIAGEN’s TissueLyser II for disruption of animal tissues to extract proteins. We suggest:

Pre-cool one 5 mm stainless steel bead and the TissueLyser II adapter at 4°C.
Chill the appropriate lysis buffer by incubating at 4°C prior to use
Freeze tissue in dry ice and immediately add 1 cold bead plus 200–300 µL of chilled lysis buffer to the TissueLyser II adapter
Disrupt twice, each time for 30 s at 20 Hz

Please be advised that these are suggestions only and have not been thoroughly tested by QIAGEN. It is the user’s responsibility to test and further optimize if necessary.

FAQ ID -9177

100 mg tissue per sample tube is the maximum amount we recommend when using the 2 x 24 Adapter Set. Larger amounts of tissue would require considered optimization, depending on the tissue type and condition.

When using 100 mg tissue, for best homogenization results, we recommend cutting the sample into 2 pieces of approximately 50 mg each prior to disruption and performing disruption twice, each time at 25 Hz for 5 min.

FAQ ID -9178

Stainless steel beads can be reused for DNA extractions with DNeasy Plant Kits. Used beads can be recovered from cell-debris and cleaned using the procedure below:

Close the cap of the 2 ml microtube and briefly vortex to dislodge the bead and pellet from the bottom of the tube. If using grinding jars for the disruption of large sample volumes, skip to step 2.
Empty the contents of the tubes/jars into a sieve and rinse the beads thoroughly with water.
Incubate beads in 0.4 M HCl for 1 min at room temperature (15–25°C) to degrade any DNA and avoid cross-contamination in future preparations.
Rinse beads thoroughly with distilled water to remove the HCl.
Dry beads before use.

You can also find these instructions in Appendix B of the DNeasy Plant Handbook.

FAQ ID -9179

Stainless steel beads sold by QIAGEN are not guaranteed to be magnetic. However, you may find some lots of the beads to be magnetic, due to small variations in the chemical compositions (which do not affect the disruptive functions of the beads).

FAQ ID -9180

We do not guarantee the stainless steel beads to be RNase free. However, when used according to our specifications for disruption of biological samples, no special treatment is needed to make the beads RNase free. This is because our chaotropic lysis buffers will usually inactivate these and RNases contained in the samples themselves.

FAQ ID -9181

QIAGEN does not carry this part. You can however purchase the O-ring from Retsch. Their part no. is 22.085.0006. Please see http://www.retsch.com/

FAQ ID -9182

Our stainless steel beads and tungsten carbide beads are ready to use.

FAQ ID -9183

The material of the grinding jar as well as the grinding ball is Teflon. The ball has a stainless steel core and a coating of Teflon.

FAQ ID -9184

Unfortunately, this is not possible. The 3 mm beads are not compatible with the 5 mm or 7 mm TissueLyser Single Bead Dispenser, as this could result in dispensation of multiple beads, and/or the dispenser may get blocked.

FAQ ID -9185

Height: 266 mm (up to 524 mm with the hood raised)

Width: 371 mm

Depth: 461 mm

Weight: Approx. 26 kg (without TissueLyser Adapter Set or Grinding Jar Set installed)

FAQ ID -9186

We do not sell 3 mm stainless steel beads for use with our TissueLyser instruments.

FAQ ID -9187

Yes, the 2 ml Sample Tubes RB (cat. no. 990381) can be used with both TissueLyser II and TissueLyser LT.

FAQ ID -9188

We do not sell ceramic beads or glass beads for use with our TissueLyser II or TissueLyser LT instruments. However, we still sell stainless steal beads and tungsten carbide beads. For more information, please go to this link.

FAQ ID -9189