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RNeasy MinElute Cleanup Kit

RNA回收和浓缩,小体积洗脱
  • 有效回收酶反应RNA
  • 可浓缩少量RNA至10 μl
  • 可回收不同方法纯化的RNA
  • 15分钟即可获得高品质RNA

RNeasy MinElute Cleanup Kit使用基于硅胶膜技术的特制RNeasy MinElute离心柱,可回收并浓缩来自于酶反应体系或其他样本中的RNA。该试剂盒还可用于RNA样本脱盐。可以将至多45 μg的RNA纯化至10 μl的体积中。整个纯化流程可在QIAcube全自动核酸纯化仪上自动完成。

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Cat No./ID: 74204
RNeasy MinElute Cleanup Kit (50)
50 RNeasy MinElute Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers

RNeasy MinElute Cleanup Kit 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。


Product Details

9
Concentration of RNA.

The indicated amounts of total RNA from HeLa cells were concentrated using the RNeasy MinElute Cleanup Kit, and 5 µl of the 12 µl eluate was used for each reaction. 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 β-actin.

9
Reliable cleanup of RNA.

The indicated amounts of total RNA from HeLa cells were concentrated using the RNeasy MinElute Cleanup Kit. Real-time RT-PCR was carried out on the LightCycler System using the QuantiTect Probe RT-PCR Kit, with primers and probes specific for the p16 gene.

9
Efficient removal of real-time RT-PCR inhibitors.
Without cleanup, inhibitors can lead to underestimation of RNA amounts in real-time quantification. Total RNA was isolated from HeLa cells using an acid-phenol method, with or without subsequent cleanup using the RNeasy MinElute Cleanup Kit. Real-time RT-PCR was carried out as described in figure "Concentration of RNA", with the indicated volumes of the RNA eluate. Apparent RNA amounts were calculated from the resulting CT values.
9
High-quality RNA.
Total RNA was isolated from HeLa cells using an acid-phenol method plus cleanup and concentration using the RNeasy MinElute Cleanup Kit. The high quality of the RNA is shown by scanning with the Agilent 2100 Bioanalyzer.
Performance
RNeasy MinElute Cleanup Kit可纯化高品质总RNA,无杂质或酶抑制物,其A 260 /A280比值为1.9–2.1(参见"High-quality RNA") 。相当于少于一个细胞的RNA的量(少至1 pg)也可以被浓缩(参见"Concentration of RNA")。可以纯化大量的RNA(至多45 µg),适用于敏感的分析(参见"Reliable cleanup of RNA")。在下游应用中可采用小量反应体积,确保提高反应效率。该试剂盒可获得高纯度的RNA,可使更多的样本用于反应中,且无抑制作用(参见"Efficient removal of real-time RT-PCR inhibitors")。
Principle

RNeasy MinElute Cleanup Kit专为从酶反应(如标记、体外转录)中纯化和浓缩RNA而设计,通过乙醇沉淀和有机提取方法分离RNA,并使RNA样本脱盐,浓缩硅胶膜操作流程制备的RNA(如,PAXgene Blood RNA制备)。RNeasy技术结合异硫氰酸胍裂解和硅胶膜纯化,简化了总RNA纯化技术。

Procedure

异硫氰酸胍裂解缓冲液和乙醇加至样本中,创造促进RNA选择性结合到RNeasy MinElute膜上的条件。然后将样本上样到RNeasy MinElute离心柱。RNA结合到硅胶膜上,污染物被高效洗去,用水洗脱得到高品质RNA。

同步回收和浓缩酶反应产物或粗RNA制备样本,可在15分钟内完成。相较而言,尤其是从小样本中,耗时的乙醇沉淀浓缩和回收会导致RNA损失。RNeasy MinElute操作流程比真空离心更快,仅离心而无需脱盐和去除其他杂质样本。RNeasy MinElute离心柱独特的设计确保将纯化得到的RNA浓缩至10 µl,用于生物芯片分析和real-time RT-PCR等下游应用。整个纯化流程可在QIAcube全自动核酸纯化仪上全自动进行。

Applications

RNeasy MinElute技术纯化得到高品质RNA,适用于各种下游应用,包括:

  • Northern印迹、点印迹和狭缝印迹
  • 终点法RT-PCR
  • 定量real-time RT-PCR
  • 芯片分析
  • Poly A+ RNA筛选

Specifications

Features
Specifications
应用 PCR, qPCR, real-time PCR, microarray
洗脱体积 10–14 µl
规格 MinElute Spin column
主要样本类型 (Crude) RNA preps
处理 Manual
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 RNA
样本量 200 µl
技术 Silica technology
每次运行或制备时间 <15 minutes
产量 45 µg

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补充实验方案 (2)
There are two protocols: follow Protocol 1 if you want to purify total RNA containing miRNA, or follow Protocol 2 if you want to purify small RNA (includes miRNA, 5S rRNA, and tRNA) and larger RNA (>200 nt) separately.
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