TissueLyser II

For medium- to high-throughput sample disruption for molecular analysis

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TissueLyser II

Cat. No. / ID: 85300

Bead mill, 100–120/220–240 V, 50/60 Hz; requires the TissueLyser Adapter Set 2 x 24 or TissueLyser Adapter Set 2 x 96 (available separately)

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The TissueLyser II is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Convenient and secure disruption process
Adapter sets optimized for high-throughput disruption
Wide range of accessories available
Reproducible results with difficult-to-lyse tissues
Front-end solution for QIAGEN automation

Product Details

The TissueLyser II simultaneously disrupts multiple biological samples through high-speed shaking in plastic tubes with stainless steel, tungsten carbide, or glass beads. Using the appropriate adapter set, up to 48 or 192 samples can be processed at the same time. Alternatively, a grinding jar set can be used to process large samples. A range of beads, bead dispensers, and collection microtubes and caps are also available.

All TissueLyser II accessories, including adapter sets, are also compatible with the TissueLyser (no longer available).

Performance

The TissueLyser II is well-suited for high-throughput disruption of human, animal, and plant tissues, bacteria, and yeast. Highly reproducible purification of high-quality DNA, RNA, miRNA, and protein is achieved, even with difficult-to-lyse tissues (see figures "Efficient disruption of animal tissues", "Reproducible RNA purification from plant tissues", "", "", "", and "").

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

Genotyping, gene expression, and proteomics applications demand effective disruption of biological samples to ensure high yields of DNA, RNA, and protein. Effortless disruption with the TissueLyser II system is achieved through high-speed shaking with beads, which beat and grind samples. The TissueLyser II system delivers thorough and rapid disruption of samples to fully release biomolecules, and also simultaneously homogenizes samples to facilitate subsequent purification procedures using QIAGEN kits (see table "QIAGEN purification kits compatible with QIAGEN disruption systems").

The TissueLyser II is an integral part of QIAGEN's complete solution for sample management — from sample collection to purification and analysis of DNA, RNA, and protein. The TissueLyser II complements QIAGEN automation for high-throughput sample preparation and analysis (see table "QIAGEN high-throughput automation"), such as the QIAsymphony SP. This easy-to-use instrument automates purification of DNA, RNA, and protein from 1–96 samples. The TissueLyser II is also compatible with QIAGEN manual sample preparation kits (see table "QIAGEN purification kits compatible with QIAGEN disruption systems").

For RNA applications, stabilization of fresh tissues in RNAprotect Tissue Reagent prevents RNA degradation during sample handling. For applications requiring purification of DNA, RNA, and protein, these 3 analytes can be immediately stabilized by placing fresh tissues in Allprotect Tissue Reagent.

QIAGEN high-throughput automation
Instrument	Purpose	Throughput
QIAsymphony SP	Purification of DNA, RNA, and protein	1–96 samples per run
BioRobot Universal System	Purification of DNA and RNA	96-well format
QIAxcel	Electophoretic analysis of DNA fragments and RNA	Up to 96 samples per run
QIAgility	Reaction setup	Up to 384 samples per run
Rotor-Gene Q	Real-time PCR and high-resolution melting (HRM) analyses	Up to 100 samples per run
PyroMark Q96 MD or ID	Methylation and mutation analyses	Up to 96 samples per run

QIAGEN purification kits compatible with QIAGEN disruption systems
Analyte purified	Sample type	QIAGEN kit
RNA	Easy-to-lyse tissue	RNeasy Kits
		RNeasy Plus Kits
		RNeasy Protect Kits
RNA	Fiber-rich tissue	RNeasy Fibrous Tissue Kits
RNA	All types of tissue	RNeasy Lipid Tissue Kits
RNA	All types of tissue	RNeasy Universal Tissue Kits
RNA	Plant tissue	RNeasy Plant Mini Kit
RNA	Yeast	RNeasy Kits
RNA	Bacteria	RNeasy Protect Bacteria Kits
microRNA	All types of tissue	miRNeasy Kits
DNA	Human tissue	QIAamp DNA Kits
DNA	Animal tissue	DNeasy Blood & Tissue Kits
Protein	Tissue	Qproteome Mammalian Protein Prep Kit
Phosphoprotein	Tissue	PhosphoProtein Purification Kit
Glycoprotein	Tissue	Qproteome Glycoprotein Kits
DNA and RNA	Tissue	AllPrep DNA/RNA Kits
DNA, RNA, and protein	Tissue	AllPrep DNA/RNA/Protein Mini Kit

Procedure

TissueLyser Adapter Sets allow processing of up to 2 x 96 samples in collection microtubes or up to 2 x 24 samples in 2 ml microcentrifuge tubes. The adapter sets can be precooled at –80°C if disrupting samples without lysis buffer. Tubes remain securely sealed during disruption to prevent cross-contamination, which is especially important for highly sensitive downstream applications, such as real-time RT-PCR or microarray analysis. Depending on the sample type, disruption is carried out using stainless steel, tungsten carbide, or glass beads. TissueLyser Bead Dispensers are available to conveniently deliver single beads into microcentrifuge tubes or to deliver 96 beads in parallel into collection microtubes. The TissueLyser II can also disrupt large samples when used in combination with Grinding Jar Sets.

Applications

The ability to process up to 192 samples per run makes the TissueLyser II the ideal front-end solution to access biological information for genomics, transcriptomics, and proteomics applications. For next-generation, high-throughput sequencing technologies, the TissueLyser II is the disruption instrument of choice. A wide range of QIAGEN sample purification kits are compatible with the TissueLyser II, and sample purification can be performed manually or can be automated using the QIAcube, QIAsymphony SP, EZ1 Advanced, or BioRobot or BioSprint workstations.

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	High-speed shaking of samples in 1.2 ml collection tubes or 2 ml microcentrifuge tubes with stainless steel or glass beads
Typical run time	15 sec - 2 x 3 minutes at 15–30 Hz
Technical data	100–120/220–240 V, 50/60Hz; variable speeds from 3 to 30 Hz (180–1800 oscillations/minute)
Kits compatible with instrument	All kits for purifying RNA/DNA/Protein
Protocol/main application on this intrument	Sample preparation/sample disruption
Throughput	2 x 96 collection microtubes (1.2 ml) or 2 x 24 microcentrifuge tubes (2ml)
Features	Convenient and secure disruption process. Adapter sets optimized for high-throughput disruption. Wide range of accessories available (e.g.,grinding jar set to process large samples). Reproducible results with difficult-to-lyse tissues. Front-end solution for QIAGEN automation.
Technology	Bead Mill

Resources

Supplementary protocol using the EZ1 Advanced DNA Investigator Large-Scale Bone Card or the EZ1 Advanced XL DNA Investigator Large-Scale Bone Card

Up to 1 x 10⁹ bacteria are disrupted and homogenized by bead-milling in a guanidine-thiocyanate-containing lysis buffer. After addition of ethanol, the sample is loaded onto an RNeasy Mini spin column. Total RNA binds to the RNeasy silica-membrane, contaminants are efficiently washed away, and high-quality RNA is eluted in RNase-free water.

This protocol has only been tested with ‘soft’ tissues (e.g., liver, spleen, thymus, heart, kidney, and brain) and may not work with ‘hard’ tissues (e.g., bone, teeth, and skin).

For high-throughput disruption of biological samples

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