Cat. No. / ID: 9002756
Cat. No. / ID: 990890
QIAGEN has tested use of the TissueRuptor II in combination with proven QIAGEN sample preparation kits. Sample disruption with the TissueRuptor II ensures maximal yields of nucleic acids and proteins that perform well in downstream applications such as PCR, western blotting and enzyme assays (see figures " Successful purification of phosphoprotein " and " High performance results in PCR"). For most tissues, disruption and homogenization using the TissueRuptor II gives comparable results to traditional rotor–stator homogenization (see figure " Pure RNA with high RIN values"). Effective disruption of tissue samples using the TissueRuptor II allows reproducible purification of high-quality DNA and RNA using QIAGEN nucleic acid purification kits (see figures " Pure RNA with high RIN values," " Reproducible purification of high-quality genomic DNA" and "Efficient disruption and homogenization of animal tissues").
Genotyping, gene expression and proteomics applications demand effective disruption of biological samples to ensure high yields of DNA, RNA and protein. The TissueRuptor 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).
The TissueRuptor 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. Optimized protocols integrate sample disruption with biomolecule purification, enabling a streamlined, efficient workflow. In addition, a range of automated solutions allow purification and analysis of biomolecules at low to high throughputs.
QIAGEN also provides RNAprotect Tissue Reagent (to stabilize RNA) and Allprotect Tissue Reagent (to stabilize DNA, RNA and protein). Tissues collected in these reagents can also be easily disrupted with the TissueRuptor II.
|Analyte purified||Sample type||QIAGEN kit|
|RNeasy Plus Kits|
|RNeasy Protect Kits|
|RNA||Fiber-rich tissue||RNeasy Fibrous Tissue Kits|
|RNA||All types of tissue||RNeasy Lipid Tissue Kits|
|RNeasy Universal Tissue Kits|
|RNA||Plant tissue||RNeasy Plant Mini Kit|
|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|
The TissueRuptor II is a portable device with a TissueRuptor Disposable Probe. The blade of the probe rotates at a high speed, causing simultaneous disruption and homogenization of a sample through a combination of turbulence and mechanical shearing. Samples that can be processed include human, animal and plant tissues (for plant tissues that are more difficult to disrupt, the TissueLyser II is recommended). Disruption at full speed for as little as 30 seconds is usually sufficient to release nucleic acids or proteins from starting material.
After disruption of a sample, the TissueRuptor Disposable Probe can be discarded, and a new probe can be used to disrupt the next sample. Time is saved and cross-contamination is avoided, as there is no need to clean and reuse the same probe. In addition, the TissueRuptor Disposable Probe is transparent, which allows visual monitoring of the sample disruption process.
The TissueRuptor II enables efficient disruption and homogenization of human, plant and animal samples, including clotted blood. Purification of RNA, DNA, total nucleic acids or protein can then be performed using QIAGEN kits.
|Disruption principle||Simultaneous disruption and homogenization by rotation of the blade of the disposable probe|
|Technology||Handheld rotor-stator homogenizer|
|Kits compatible with instrument||All Kits for purification of RNA/DNA/protein|
|Technical data||AUS/EU/UK: 230 V, 144 W, 50/60 Hz, 5000-35,000 rpm;<br /> NA: 120 V, 144 W, 50/60 Hz, 5000-35,000 rpm;<br /> Japan: 100 V, 125 W, 50/60 Hz, 5000-28,000 rpm|
|Typical run time||15–120 seconds|
|Features||Rapid, effective disruption of a range of sample types. Disposable probes minimize risk of cross-contamination. Time savings through use of disposable probes. Visual monitoring of disruption using transparent probes. Seamless integration with QIAGEN sample technologies.|
|Throughput||1 sample per run|
Yield and integrity of genomic DNA depend on the disruption and homogenization method used with the Allprep DNA/RNA Kits.
Homogenization with the TissueRuptor (or other rotor–stator homogenizer, such as the Polytron) or the TissueLyser results in greater DNA fragmentation (depending on homogenization time and intensity). However, shorter DNA fragments are easier to elute, so DNA yields will be higher.
In contast, homogenization using the QIAshredder (or a syringe and needle) is more gentle, resulting in less DNA fragmentation. However, longer DNA fragments are more difficult to elute, so DNA yields may be lower.
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.
Since tissues incubated in Allprotect Tissue Reagent are harder than fresh tissues, longer homogenization times might be necessary (for details, see the Allprotect Tissue Reagent Handbook).
Furthermore, we do not recommend using rotor–stator homogenizers with a metal probe (e.g., Polytron) if proteins will be purified subsequently, as the metal probe can heat the sample.