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Kits for RNA isolation from tissue Kits for RNA isolation from all sample types

Format:	Mini spin columns
Sample sources:	Whole blood and tissue
Sample size:	50 µl – 1.5 ml
Preparation time:	<1 hour
Typical yield:	1–5 µg RNA per ml healthy blood or up to 100 µg RNA from tissue
Elution volume:	30–100 µl

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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.

Procedure

The QIAamp RNA Blood Mini Kit simplifies isolation of RNA from blood with a fast spin-column procedure (see figure "QIAamp RNA Blood Mini Procedure"). No phenol–chloroform extraction is required. RNA binds specifically to the QIAamp silica-gel membrane while contaminants pass through. PCR inhibitors such as divalent cations and proteins are completely removed in two efficient wash steps, leaving pure RNA to be eluted in either water or a buffer provided with the kit.

QIAamp technology yields total cellular RNA from fresh whole blood and other sample sources ready to use in RT-PCR and blotting procedures. QIAamp sample preparation technology is fully licensed, allowing QIAamp purified nucleic acids to be used in any molecular assay or other downstream application without risk of patent infringement.

Procedure

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 30–100 µl RNase-free water (provided with the kit) for direct use in any downstream application.

Downstream applications

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").

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