For isolation of cellular RNA from fresh whole blood
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| Format: | Mini spin columns |
| Sample sources: | Whole blood and tissue |
| Sample size: | 50 µl 每 1.5 ml |
| Preparation time: | <1 hour |
| Typical yield: | 1每5 µg RNA per ml healthy blood or up to 100 µg RNA from tissue |
| Elution volume: | 30每100 µl |
The
QIAamp RNA Blood Mini Kit is designed for isolation of cellular RNA from up to
1.5 ml of fresh, whole human blood stabilized with any common anticoagulant,
such as citrate, heparin, or EDTA. In addition, total cellular RNA can be
isolated from tissue samples.
The QIAamp procedure completely removes RNases, contaminants, and enzyme inhibitors, yielding high-quality RNA suitable for any downstream application (see figures "High-Quality RNA for Northern Analysis" and "Reliable RT-PCR Analysis"). Red blood cells are selectively lysed and white cells collected by centrifugation. White cells are then lysed using highly denaturing conditions which immediately inactivate RNases. After homogenization using the QIAshredder spin column, the sample is applied to the QIAamp spin column. Total RNA binds to the QIAamp membrane and contaminants are washed away, leaving pure RNA to be eluted in 30每100 µl RNase-free water (provided with the kit) for direct use in any downstream application.
High-Quality RNA for Northern 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). 40 µl of a 60 µl eluate were loaded per lane. M: 0.24每9.5 kb RNA ladder.
The QIAamp RNA Blood Mini Kit provides the highest-quality RNA with minimum copurification of DNA. However, as with any RNA purification method, some DNA contamination can be expected. For certain RNA applications that are sensitive to very small amounts of DNA, it may be necessary to remove any remaining DNA. In these cases, the QIAGEN RNase-Free DNase Set provides convenient on-column DNase treatment of RNA samples during QIAamp RNA procedures.
For purification of RNA from cell-free body fluids, the QIAamp Viral RNA Mini Kit should be used; for purification of RNA from tissues, QIAGEN also offers the RNeasy system. See also the QIAamp Selection Guide.
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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. 每 PCR was performed as above but eluate was replaced with distilled water; + positive control cDNA fragment. M: 100 bp DNA ladder. |
紱釬忒聊: QIAamp RNA Blood Mini Handbook (PDF version, 209KB)