从新鲜的全血样本中纯化胞内RNA

  • 快速纯化高品质的即用型RNA
  • 无需有机提取或乙醇沉淀
  • 重复性好,产量高
  • 完全去除污染物和抑制剂

QIAamp RNA Blood Mini Kit使用硅胶膜,可从多至1.5 ml的人类新鲜全血中纯化胞内RNA。可以使用常规的抗凝剂如柠檬酸盐、肝素或EDTA等稳定血液样本。使用QIAshredder离心柱对样本进行均化处理后进行快速离心,从而简化了纯化RNA的流程。可在QIAcube全自动核酸纯化仪上全自动完成纯化处理。

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QIAamp RNA Blood Mini Kit (50)
For 50 RNA preps: 50 QIAamp Mini Spin Columns, 50 QIAshredder Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free reagents and buffers
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The QIAamp RNA Blood Mini 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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QIAamp RNA Blood Mini procedure.|High-quality RNA for northern analysis.|Reliable RT-PCR analysis.|
|Formaldehyde agarose gel and corresponding northern blot (GAPDH-probed) of total RNA isolated with the QIAamp RNA Blood Mini Kit from 0.5, 1.0, and 1.5 ml (left to right) of healthy whole human blood (with indicated anticoagulants). Forty microliters of a 60 µl eluate were loaded per lane. M: 0.24–9.5 kb RNA ladder.|RT-PCR of total RNA from blood. Total RNA from 50 µl of healthy whole human blood (with indicated anticoagulants) was purified using the QIAamp RNA Blood Mini Kit and eluted with 50 µl of RNase-free water. For each sample, three RT-PCR reactions were performed (left to right) using 5 µl or 15 µl of the eluate or a control (15 µl eluate, RT omitted). For RT-PCR, samples were digested with RNase-free DNase and reverse transcription performed using an oligo-dT primer. 1/10 (2.5 µl) of the cDNA was amplified using GAPDH primers. 1/5 of the PCR reaction was loaded per lane.  PCR was performed as above but eluate was replaced with distilled water; + positive control cDNA fragment. M: 100 bp DNA ladder.|
Performance

QIAamp纯化流程可完全去除RNases、污染物和酶抑制剂,制备的高品质RNA适用于多种下游应用(参见"High-quality RNA for northern analysis"和"Reliable RT-PCR analysis")。

QIAamp RNA Blood Mini Kit制备的RNA品质高,含有最少量的DNA。RNA的纯化不可避免存在DNA污染。但对于某些痕量DNA敏感的RNA应用则必须去除残留的DNA,对于这些应用,QIAGEN RNase-Free DNase Set(可选购)为QIAamp RNA纯化流程提供简便的柱上DNase酶处理操作,去除残留DNA。

Principle

QIAamp RNA Blood Mini Kit使用硅胶膜技术纯化细胞RNA,无需苯酚-氯仿抽提。RNA特异性结合到QIAamp硅胶膜上,污染物流出。通过两步高效洗涤完全去除二价阳离子和蛋白质等PCR抑制剂,再用水或试剂盒中的缓冲液洗脱结合在离心柱上的纯DNA。

QIAamp技术从新鲜的全血样本或其他样本来源中纯化的总细胞RNA,可即时用于PCR和印迹实验中。QIAamp样本制备技术完全得到验证认可。

Procedure
QIAamp RNA Blood Mini Kit使用快速离心操作简化从血液中纯化RNA的操作流程(参见"Procedure")。选择性裂解红细胞,离心收集白细胞。在高度变性条件下白细胞裂解,RNases很快失活。使用QIAshredder离心柱均质化后,将样本上样到QIAamp离心柱。总RNA结合到QIAamp膜,污染物流出,在30–100 µl的无RNase水中洗脱残留的纯RNA(试剂盒提供),可即时用于多种下游应用。
Applications

QIAamp RNA Blood Mini Kit可从多至1.5 ml的新鲜人体全血中纯化细胞RNA。可以使用常规的抗凝剂如柠檬酸盐、肝素或EDTA等稳定血液样本。也可从组织样本中纯化得到总细胞RNA。

Feature
Specifications
Applications PCR, real-time PCR, microarray
Elution volume 30–100 µl
Format Spin columns
Main sample type Whole blood, tissue
Processing Manual (centrifugation)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Cellular RNA
Sample amount 50–1.5 ml
Technology Silica technology
Time per run or per prep <1 hour
Yield 1–5 µg

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Kit Handbooks
1
For total RNA purification from human whole blood
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