For purification of up to 45 µg total RNA from cell and tissue samples
- Fast procedure delivering high-quality total RNA in minutes
- Ready-to-use RNA for high performance in any downstream application
- Consistent RNA yields from very small amounts of starting material
- No phenol/chloroform extraction
- No CsCl gradients, no LiCl or ethanol precipitation
The RNeasy Micro Kit is designed for purification of up to 45 μg RNA from small cell and tissue samples. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. RNA purification using the RNeasy Micro Kit can be fully automated on the QIAcube Connect. For larger samples, the RNeasy Mini Kit (spin-column binding capacity of 100 µg RNA), RNeasy Midi Kit (spin-column binding capacity of 1 mg RNA), and RNeasy Maxi Kit (spin-column binding capacity of 6 mg RNA) are also available.
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RNeasy Micro Kit (50)
50 RNeasy MinElute Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free DNase I, Carrier RNA, RNase-free Reagents and Buffers
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74004
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The RNeasy Micro Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
产量稳定,适用于敏感应用。|从活检针中纯化获得高品质总RNA。|RNeasy Micro实验流程。|高效的柱上基因组DNA去除。|从单细胞中可靠纯化RNA。|
5位不同的用户使用RNeasy Micro Kit,从2 mg图示大鼠组织中分离总RNA,重复两次。在ABI Sequence Detection System上使用QuantiTect Probe RT-PCR Kit及c-jun特异性引物和探针进行定量real-time RT-PCR。|使用19.5口径的活检针从兔肝脏中进行细针抽吸活检,样本立即置于RNAlater TissueProtect Tube内稳定。使用RNeasy Micro Kit分离总RNA。采用Agilent 2100 Bioanalyzer分析显示了高质量的RNA。|RNeasy Micro Kit可用于从较小样本中分离总RNA。优化的实验方案适用于显微切割组织、少量标准组织、富含纤维的组织及细胞的RNA分离。还可提供RNA回收和浓缩实验方案。|使用RNeasy Micro Kit从图示量的HeLa细胞中分离总RNA。在ABI Sequence Detection System上使用QuantiTect Probe PCR Kit及c-jun特异性引物和探针进行定量real-time PCR。在逆转录后(+RT)以及未经逆转录步骤直接(-RT)扩增各样本,以评估柱上DNA酶切效率。"-RT"样本中无信号表示未检测到DNA。在"-RT"结果中,仅有50,000个细胞数据点可见。|使用RNeasy Micro Kit从图示量的HeLa细胞中分离总RNA。[A] 扩增曲线。[B] 交叉点(CP)值与细胞数的相关性。在LightCycler System上使用QuantiTect Probe RT-PCR Kit及β-actin特异性引物和探针进行real-time RT-PCR。|
原理
The RNeasy Micro Kit is designed for isolation of total RNA from small 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 by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification and the unique design of RNeasy MinElute spin columns enables minimal elution volumes, as little as 10 µl, to concentrate the RNA. The small elution volume means that reaction volumes can be kept small in downstream applications, giving increased reaction efficiency. The high purity allows the complete sample to be used in downstream reactions, such as reverse transcription, without enzyme inhibition.
操作流程
The RNeasy Micro Kit is designed for isolation of total RNA from small samples. Samples (<5 mg tissue, or <5 x 105 cells) 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 MinElute spin column and RNA (up to 45 µg) binds to the silica membrane. RNase-free DNase, provided in the RNeasy Micro Kit, enables simple and efficient on-column digestion of genomic DNA for sensitive applications. Then DNase and any contaminants are efficiently washed away, and pure, concentrated RNA is eluted in 10–14 µl water.
应用
Highly reproducible isolation of RNA in a small elution volume (10 µl) using the RNeasy Micro Kit makes the highly pure RNA well-suited for sensitive quantitative gene expression analyses, including real-time RT-PCR starting with as little as one cell.
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Feature
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Specifications
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Applications
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Northern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
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Elution volume
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10–14 µl
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Format
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Spin column
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Main sample type
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Tissue, cells
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Processing
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Manual
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Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
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RNA
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Sample amount
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<5 mg
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Technology
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Silica technology
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Time per run or per prep
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30–40 minutes
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Yield
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Varies
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FAQ ID - 3388
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I received one of the kits in the list below containing the MinElute columns, however they were left out for a while and not stored at 2-8°C upon receipt. Can I still use them? AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro
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FAQ ID -942
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For purification of total RNA from small samples, including
- animal and human cells
- animal and human tissues
- fibrous tissues
- microdissected cryosections
and for RNA cleanup and concentration
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Show details
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Images
Highly reproducible yields for sensitive applications.
Total RNA was isolated by 5 different users in duplicate from 2 mg of the indicated rat tissues, using the RNeasy Micro Kit. Quantitative, real-time RT-PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for c-jun.
High-quality total RNA from fine needle aspirates.
A fine needle aspirate was obtained from rabbit liver using a 19.5-gauge biopsy needle, and the sample was immediately stabilized in an RNAprotect Tissue Tube. Total RNA was isolated using the RNeasy Micro Kit. The high quality of the RNA is shown by scanning with the Agilent 2100 Bioanalyzer.
RNeasy Micro procedure.
The RNeasy Micro Kit is designed for isolation of total RNA from small samples. Optimized protocols are provided for RNA isolation from microdissected tissues, small amounts of standard tissues, fiber-rich tissues, and cells. Protocols are also available for RNA cleanup and concentration.
Efficient on-column removal of genomic DNA.
Total RNA was isolated from the indicated number of HeLa cells using the RNeasy Micro Kit. Quantitative, real-time PCR was carried out on an ABI Sequence Detection System using the QuantiTect Probe PCR Kit, with primers and probes specific for c-jun. In order to evaluate the efficiency of the on-column DNase digestion, each sample was amplified after reverse transcription (+RT) and directly, without a reverse transcription step (-RT). The lack of signal in the "-RT" samples indicates that no DNA was detectable. Of the "-RT" results, only the 50,000 cell data points are visible.
Reliable RNA isolation from a single cell.
Total RNA was isolated from the indicated number of HeLa cells using the RNeasy Micro Kit. [A] Amplification plot. [B] Correlation of crossing point (CP) values and cell numbers. Real-time RT-PCR was carried out on the LightCycler System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for β-actin.
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