For medium- to high-throughput sample disruption for molecular analysis
- 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
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 accessories for the TissueLyser II, including adapter sets, are also compatible with the TissueLyser (no longer available).
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TissueLyser II
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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85300
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TissueLyser II适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。
Reliable detection of miRNAs.|Efficient disruption of animal tissues.|Reproducible RNA purification from plant tissues.|Intact protein suitable for all types of analysis.|High-quality DNA from animal tissues.|High-quality DNA from plant tissues.|TissueLyser II.|
Rat kidney was stabilized in RNAprotect Tissue Reagent, and 25 mg samples were disrupted in QIAzol Lysis Reagent for 2 x 5 minutes at 25 Hz using the TissueLyser II. Total RNA containing miRNA was purified using the miRNeasy Mini Kit, using either the manual procedure (Manual) or the automated procedure with the QIAcube (QIAcube). Real-time RT-PCR using the miScript System was carried out to detect 2 different miRNAs, miR-16 and miR-25.|Tissues (20 mg) were either frozen (Frozen) or stabilized in RNAlater RNA Stabilization Reagent (RNAlater), and then disrupted using the TissueLyser II (2 x 2 minutes for liver and muscle; 2 x 5 minutes for skin). RNA was purified using the RNeasy Mini Kit (liver), RNeasy Fibrous Tissue Mini Kit (skin), or RNeasy Lipid Tissue Mini Kit (muscle). [A] Analysis on a 1.2 % formaldehyde agarose gel shows sharp ribosomal RNA bands, indicating intact RNA. [B] RNA yields were quantified by A260 nm absorbance measurements.|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).|Various rat tissues (25 mg each)
stabilized in Allprotect Tissue Reagent were disrupted using the TissueLyser LT or TissueLyser II.
Total protein was purified using the Qproteome Mammalian Protein Prep Kit and then
analyzed by western blotting. Transfer membrane stained with Ponceau S, and western
blot. LT: TissueLyser LT; II: TissueLyser II; M: markers.|Various rat
tissues (25 mg each) stabilized in Allprotect Tissue Reagent
were disrupted using the TissueLyser LT or TissueLyser II.
DNA was purified on the QIAcube using the DNeasy Blood
& Tissue Kit, and then used in PCR with the HotStarTaq Plus
Master Mix Kit and a PGK1 primer system. The 120 bp PCR
product was analyzed on the QIAxcel using the QIAxcel
DNA High Resolution Kit. LT: TissueLyser LT; II: TissueLyser II;
M: markers.|Various
plant tissues (100 mg each) were disrupted in precooled
adapters using the TissueLyser LT or TissueLyser II. DNA was
purified on on the QIAcube using the DNeasy Plant Mini Kit
and then analyzed on an agarose gel. LT: TissueLyser LT;
II: TissueLyser II; M: markers.||
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", " Reproducible DNA purification from animal tissue", " Purification of intact protein from animal tissue", " Reproducible RNA purification from Gram-positive bacteria", and " 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.
| 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 |
| RNA |
Easy-to-lyse tissue |
RNeasy Kits
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| 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 |
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.
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特点
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参数
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破碎原理
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High-speed shaking of samples in 1.2 ml collection tubes or 2 ml microcentrifuge tubes with stainless steel or glass beads
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特点
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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.
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与仪器兼容的试剂盒
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All kits for purifying RNA/DNA/Protein
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该仪器的实验方案/主要应用
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Sample preparation/sample disruption
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技术参数
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100–120/220–240 V, 50/60Hz; variable speeds from 3 to 30 Hz (180–1800 oscillations/minute)
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技术
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Bead Mill
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通量
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2 x 96 collection microtubes (1.2 ml) or 2 x 24 microcentrifuge tubes (2ml)
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常规运行时间
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15 sec - 2 x 3 minutes at 15–30 Hz
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For high-throughput disruption of biological samples
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Supplementary protocol using the EZ1 Advanced DNA Investigator Large-Scale Bone Card or the EZ1 Advanced XL DNA Investigator Large-Scale Bone Card
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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.
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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).
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图片
可靠检测miRNA。
将大鼠肾脏置于RNAlater RNA Stabilization Reagent中稳定,使用TissueLyser II,在QIAzol Lysis Reagent中25 Hz破碎25 mg样本2 x 5分钟。使用miRNeasy Mini Kit,采用手动步骤 (Manual) 或在QIAcube全自动核酸纯化仪 (QIAcube) 上进行自动化处理,纯化包含miRNA的总RNA。使用miScript System进行Real-time RT-PCR,检测2种不同的miRNA——miR-16和miR-25。
高效破碎动物组织。
冷冻 (Frozen) 组织 (20 mg) 或将其置于RNAlater RNA Stabilization Reagent (RNAlater) 中稳定,然后使用TissueLyser II破碎 (肝脏和肌肉破碎2 x 2分钟;皮肤破碎2 x 5分钟)。使用RNeasy Mini Kit (肝脏)、RNeasy Fibrous Tissue Mini Kit (皮肤) 或RNeasy Lipid Tissue Mini Kit (肌肉) 纯化RNA。[A] 采用1.2 %的甲醛琼脂糖凝胶分析显示了清晰的核糖体RNA条带,表示具有完整的RNA。[B] 利用A260 nm吸光度测量,定量RNA产量。
从植物组织中纯化RNA具有可重复性。
使用TissueLyser II破碎冷冻的植物叶子 (2 x 1分钟)。使用RNeasy Plant Mini Kit纯化RNA,并在1.2 %的甲醛琼脂糖凝胶上进行分析。核糖体RNA条带清晰均匀,表示可重复纯化出完整的RNA。T:番茄 (100 mg);A:拟南芥 (25 mg);C:棉花 (100 mg);M:玉米 (100 mg);R:油菜 (100 mg)。
从动物组织中纯化DNA,具有可重复性。
将一只大鼠的心脏切成96 x 25 mg的小块,然后使用TissueLyser II破碎 (2 x 15秒)。在QIAcube全自动核酸纯化仪上使用DNeasy Blood & Tissue Kit (采用Reagent DX最大程度减少裂解起泡现象) 进行自动DNA纯化。48个样本的琼脂糖凝胶分析显示了各样本中的DNA量一致。M:分子量标准。
从动物组织中纯化完整的蛋白。
将组织样本(50 mg) 置于Mammalian Cell Lysis Buffer中预冷至4°C,使用TissueLyser II 20 Hz破碎3 x 90秒。使用针对∝-tubulin、β-actin和GAPDH的抗体对裂解物 (15 μg) 进行western印迹分析。同时分析纯化的蛋白质 (1 μg、100 ng和10 ng),作为阳性对照。Lu:肺;He:心脏;Sp:脾;Li;肝;Ki:肾;Br:脑;Th:胸腺。
从革兰氏阳性菌中纯化RNA,具有可重复性。
使用RNAprotect Bacteria Reagent稳定B. subtilis培养物。使用TissueLyser II,在裂解缓冲液中30 Hz破碎单个样本 (各2.5 x 108个细胞) 5分钟。使用RNeasy Protect Bacteria Mini Kit纯化总RNA,并在甲醛琼脂糖凝胶上进行分析。
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