- 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
- 微量起始样本, RNA产量稳定
- 无需酚/氯仿抽提
- 无需CsCl梯度离心,无需LiCl或乙醇沉淀
RNeasy Micro Kit专用于从微量细胞和组织样本中纯化至多45 μg RNA。使用RNAlater RNA Stabilization Reagent或Allprotect Tissue Reagent方便的稳定组织样本,使用TissueRuptor或TissueLyser体系高效破碎样本。可在QIAcube全自动核酸分析仪上应用RNeasy Micro Kit实现RNA纯化全自动化。处理更大的样本,可使用RNeasy Mini Kit(离心柱结合能力为100 µg RNA)、RNeasy Midi Kit(离心柱结合能力为1 mg RNA)或RNeasy Maxi Kit(离心柱结合能力为6 mg)。
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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
RNeasy Micro Kit专用于从微量样本中纯化总RNA,如显微切割组织、细针穿刺样本、微量纤维组织包括心脏和肌肉组织、微量细胞、甚至是单细胞(如FACS筛选细胞)。处理显微切割FFPE组织,我们推荐使用RNeasy FFPE Kit。RNeasy技术整合了苯酚/异硫氰酸胍裂解的严谨性和硅胶膜纯化的速度和纯度,简化了总RNA的纯化技术。RNeasy MinElute离心柱独特的设计可确保最小的洗脱体积,小至10 µl,实现RNA的高度浓缩。小洗脱体积意味着下游的反应可控制在小体积范围内,提高反应效率。高纯度确保全部样本可用于反转录等下游应用,无酶抑制。
操作流程
RNeasy Micro Kit专用于从微量样本中纯化总RNA。先对样本(<5 mg组织或<5 x 105个细胞)进行破碎和匀浆。在裂解物中加入乙醇以提供合适的结合条件。然后将裂解物上样至RNeasy MinElute离心柱,RNA(至多45 µg)结合到硅胶膜上。RNeasy Micro Kit包含RNase-free DNase,确保简单、高效的柱上基因组DNA消化,适用于敏感的应用。DNase和其他污染物被高效洗去后,10–14 µl水洗脱得到高浓度的纯化RNA。
应用
RNeasy Micro Kit纯化得到小洗脱体积(10 µl)RNA,重复性好,适用于敏感的定量基因表达分析,包括以单细胞为起始样本的real-time RT-PCR。
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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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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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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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