从细胞或组织样本中纯化至多45 μg的总RNA

  • 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
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.
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Highly reproducible yields for sensitive applications.|High-quality total RNA from fine needle aspirates.|RNeasy Micro procedure.|Efficient on-column removal of genomic DNA.|Reliable RNA isolation from a single cell.|
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.|A fine needle aspirate was obtained from rabbit liver using a 19.5-gauge biopsy needle, and the sample was immediately stabilized in an RNAlater TissueProtect 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.|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.|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.|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.|
Performance
RNeasy Micro Kit可从微量样本中得到高产量总RNA,且重复性好。从少量细胞,包括单细胞以及微量标准组织样本中都能纯化得到总RNA(参见"Reliable RNA isolation from a single cell"、"Highly reproducible yields for sensitive applications"和"High-quality total RNA from fine needle aspirates")。RNeasy Micro Kit通过高效的柱上基因组DNA消化可使珍贵的样本在定量基因表达分析中得到最大的敏感度,如real-time RT-PCR(参见"Efficient on-column removal of genomic DNA")。
Principle

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。

Procedure

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。

Applications

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。

Feature
Specifications
Applications Northern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
Elution volume 10–14 µl
Format Spin column
Main sample type Tissue, cells
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount <5 mg
Technology Silica technology
Time per run or per prep 30–40 minutes
Yield Varies

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Kit Handbooks
1
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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