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分子生物学
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细胞生物学
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蛋白质组学
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蛋白凝胶纯化
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生化试剂
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仪器设备
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通用实验耗材
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色谱耗材
特点
- · 数分钟内快速纯化得到高品质总RNA
- · 即用型RNA在各种下游应用中表现良好
- · 大量起始样本，RNA产量稳定
- · 无需酚/氯仿抽提
- · 无需CsCl梯度离心，无LiCl或者乙醇沉淀
产品详情
RNeasy Kits 是总RNA分离的金标准。它们使用硅胶膜RNeasy spin columns 或96孔板从少量到大量细胞、组织和酵母中快速纯化高质量RNA。使用RNAprotect Tissue Reagent 或 Allprotect Tissue Reagent 可以方便地稳定组织样品，并使用TissueRuptor II或TissueLyser II或LT系统有效破坏组织样品。RNeasy 96 Kit能够使用硅膜RNeasy 96板从多达96个培养细胞样品中高通量纯化总RNA。专用RNeasy QIAcube Kit 可在QiaCube Connect上自动纯化1-12个样品。
性能
RNeasy Kits deliver highly reproducible yields of total RNA from small to large samples. Total RNA can reliably be purified from small numbers of cells, including a single cell, as well as from small amounts of standard tissues (参见 "Reliable RNA isolation from a single cell", "Highly reproducible yields for sensitive applications" and "High-quality total RNA from fine needle aspirates") using the RNeasy Micro Kit. RNA purified using the RNeasy Micro Kit provides maximum sensitivity with precious samples in quantitative gene expression analyses, such as real-time RT-PCR, by efficient on-column digestion of genomic DNA (参见 "Efficient on-column removal of genomic DNA").
Total RNA is easily purified from animal or human cells, animal or human tissues, and yeast (see table “Total RNA yields obtained with RNeasy Kits” and figure "RT-PCR of RNA from as few as 100 cells").
使用RNeasy Kits 获得的总RNA产量
| Source | Starting material | Average yield | ||||||||
| Animal cells | Micro | Mini | Midi | Maxi | 96-well | Micro | Mini | Midi | Maxi | c |
| LMH | 5 x 105 | 1 x 106 | 7 x 107 | 5 x 108 | 1 x 105 | – | 12 µg | 850 µg | 5.7 mg | 1.3 µg* |
| HeLa | 5 x 105 | 1 x 106 | 7 x 107 | 4 x 108 | 1 x 105 | 5mg | 15 µg | 1000 µg | 6.0 mg | 1.6 µg* |
| COS-7 | 5 x 105 | 1 x 106 | 3 x 107 | 1.8 x 108 | 1 x 105 | 17.5 mg | 35 µg | 950 µg | 5.8 mg | 3.1 µg* |
| Lymphocytes (unstimulated) | 5 x 105 | 1 x 106 | 1 x 108 | 5 x 108 | – | – | 0.5 µg | 50 µg | 0.3 mg | – |
| Huh7 | 5 x 105 | – | – | – | 1 x 105 | 7.5mg | – | – | – | 2.0 µg* |
| Jurkat | 5 x 105 | – | – | – | 1 x 105 | – | – | – | – | 1.4 µg* |
| Mouse tissue | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Liver | 5 mg | 10 mg | 200 mg | 1 g | – | 15 µg | 40 µg | 700 µg | 3.6 mg | – |
| Lung | 5 mg | 10 mg | 100 mg | 0.5 g | – | 5 µg | 10 µg | 130 µg | 0.6 mg | – |
| Spleen | 5 mg | 10 mg | 200 mg | 1 g | – | 15 µg | 35 µg | 600 µg | 3.2 mg | – |
| Yeast cells | ||||||||||
| S. cerevisiae | – | 1 x 107 | 2 x 108 | 1 x 109 | – | – | 25 µg | 450 µg | 2.4 mg | – |
*数量可能因发育阶段、使用的生长条件等而异。
The dedicated RNeasy Mini QIAcube Kit, including rotor adapters preloaded with RNeasy spin columns and elution tubes, delivers greater convenience and time savings (参见 "Significant time savings with dedicated QIAcube Kits").
Total RNA purified with the RNeasy Maxi Kit is of high quality and is suitable for many downstream applications (参见 "High-quality RNA from a variety of samples"). Total RNA is easily purified with the RNeasy Maxi Kit from large amounts of starting material including animal or human cells, animal or human tissues, and yeast cells (see table “Total RNA yields obtained with RNeasy Kits”).
The RNeasy 96 system provides fast and reproducible total RNA purification for high-throughput gene expression profiling. The RNA is suitable for sensitive applications such as quantitative, real-time RT-PCR and microarray analysis (参见 "High-quality RNA for sensitive analysis of a low-copy transcript"). Sample sizes range from 10 to 5 x 105 cells (see figure "RT-PCR of RNA from as few as 100 cells"), and high-quality RNA can be purified from large numbers of samples (参见 "Reproducible yields of high-quality RNA"). Individual variations are low throughout the entire purification process; TaqMan® threshold-cycle values are easily achieved at the end of the process with a coefficient of variation (CV) less than 3% using the QuantiTect Probe RT-PCR Kit (参见 "Repeatability of fully automated RNA purification").
原理
RNeasy Kits allow efficient purification of total RNA from small to large amounts of starting material, including samples such as microdissected tissues and fine needle aspirates, from milligram amounts of fibrous tissues including heart and muscle tissue, and from small numbers of cells down to single cells (e.g., FACS sorted cells). For microdissected FFPE tissues, we recommend the RNeasy FFPE Kit. RNeasy technology simplifies total RNA isolation from cells, tissues and yeast by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification. RNeasy Kits provide the highest-quality RNA with minimum copurification of DNA.
步骤
RNeasy technology simplifies total RNA isolation (参见 “Amount of starting material for RNeasy procedures”). Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the RNeasy silica membrane and RNA binds to the silica membrane, and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in water. A variety of special application protocols is also available.
Automated processing on the QIAcube Connect
The QIAcube Connect uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated, low-throughput sample prep into laboratory workflows. All steps in the purification procedure are fully automated — and up to 12 samples can be processed per run. The QIAcube Connect together with the dedicated RNeasy Mini QIAcube Kit provides fast, easy, and convenient RNA purification.
High-throughput processing in 96-well format
The RNeasy 96 system provides a fast and efficient procedure. Cells can be grown and directly lysed in 96-well cell-culture dishes. After transfer of the lysates to the wells of the RNeasy 96 plate, RNA binds to the silica membrane in each well, and contaminants are washed away. Pure RNA is then eluted in RNase-free water into individual collection tubes suitable for long-term storage and is ready to use for any experiment. The RNeasy 96 Kit includes protocols for total RNA isolation, cytoplasmic RNA isolation, and RNA cleanup. RNeasy 96 plates can be processed on the QIAvac 96 vacuum manifold and/or the 96-Well-Plate Centrifuge System.
RNeasy程序的起始材料量
| Amount of starting material | RNeasy Micro Kit | RNeasy Mini Kit | RNeasy Mini QIAcube Kit | RNeasy Midi Kit | RNeasy Maxi Kit | RNeasy 96 Kit |
| Cells | <5 x 105 cells | 10 to 1 x 107 animal or human cells | 1 x 107 cells | 5 x 106 to 1 x 108 animal or human cells | Up to 5 x 108 animal or human cells | 10–5 x 105 cells |
| Tissue | <5 mg tissue | 0.5–30 mg animal or human tissues | 30 mg tissue | 20–250 mg animal or human tissues | 150 mg to 1 g tissue | – |
| Yeast | <5 x 107 yeast cells | – | 2 x 107 to 5 x 108 yeast cells | 2.5 x 109 yeast cells | – |
应用
使用RNeasy技术纯化的RNA的A260/280比率为1.9–2.1（以10mM Tris·Cl、pH 7.5测量），非常适合用于所有应用。下游应用包括：
· 转录组测序
· 定量实时RT-PCR
· 从一个细胞开始的实时RT-PCR
· 终点RT-PCR
· Northern、点和槽印迹法
· 阵列分析
· Poly A+RNA选择
RNeasy试剂盒的比较
| Features | RNeasy Micro Kit | RNeasy Mini Kit | RNeasy Mini QIAcube Kit | RNeasy Midi Kit | RNeasy Maxi Kit | RNeasy 96 Kit |
| Applications | RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, Microarray analysis | RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, Microarray analysis | RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, Microarray analysis | RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, Microarray analysis | RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, Microarray analysis | RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, Microarray analysis |
| Elution volume | 10–14 µl | 30–100 µl | 30–100 µl | 300–500 µl | 800–2400 µl | 45–140 µl |
| Format | Spin column | Spin column | Spin column, preloaded into rotor adapters | Spin column | Spin column | 96-well plate |
| Main sample type | Tissue, cells | Tissue, cells, yeast | Tissue, cells, yeast | Tissue, cells, yeast | Tissue, cells, yeast | Cultured cells |
| Processing | Manual (centrifugation) | Manual (centrifugation) | Automated (QIAcube Connect) | Manual (centrifugation) | Manual (centrifugation) | Manual (centrifugation or vacuum) |
| Purification of total RNA, miRNA, | Total RNA | Total RNA | Total RNA | Total RNA | Total RNA | Total RNA |
| Sample amount | <5 mg tissue, <5 x 105 cells | 0.5–30 mg tissue, 10 to 1 x 107 animal or human cells | 0.5–30 mg tissue, 1 x 107 cells | 20–250 mg tissue, 5 x 106 to 1 x 108 animal or human cells | 150 mg – 1 g tissue, Up to 5 x 108 animal or human cells | 5 x 105 |
| Technology | Silica technology | Silica technology | Silica technology | Silica technology | Silica technology | Silica technology |
| Time per run or per prep | 30–40 minutes | 20 minutes | – | <1 hour | 1 hour | 30–40 minutes |
| Yield | Varies | Varies | Varies | Varies | Varies | 1.3–3.1 µg |
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