|Format||Mini spin columns|
|Sample material||≤200 µl phenol/chloroform extract, reaction mixture, or <105 cells|
|Fragment size||>200 nt|
|Typical RIN (RNA integrity number)||Depends on sample quality (no significant loss of RIN detected)|
|Elution volume||40–120 µl|
|Preparation time||20 min/6 preps|
|Binding capacity||200 µg|
NucleoSpin RNA Clean-up
- Complete removal of RT-PCR inhibitors
- Time-saving procedure based on NucleoSpin RNA, without DNase digestions and homogenization steps
- RNA cleanup from pre-purified RNA (phenol/chloroform), enzymatic reactions (e.g., amplification reactions, labeling reactions)
- For isolation of RNA from cells and tissue with minimal DNA contamination, we recommend rDNase-containing NucleoSpin RNA kits
RNA clean-up for:
- Pre-purified RNA (e.g., Trizol)
- Reaction mixtures
- Biotinylated RNA
- RNA isolation from up to 105 cultured cells (whenever co-purification of some genomic DNA is acceptable, kit does not contain rDNase)
- Typical downstream applications: enzymatic-labeling reactions, RT-PCR, DNA/RNA-based chip hybridizations
NucleoSpin RNA Clean-Up kits are recommended for the cleanup of total RNA from RNA preparations which contain unacceptable amounts of RT-PCR inhibitors, such as RNA prepared with phenol-chloroform based methods. These kits are also suitable for the isolation of RNA from small amounts of cultured cells whenever copurification of some genomic DNA is acceptable. The kits allow purification of pure RNA with an A260/A280 ratio generally exceeding 1.9 (measured in TE buffer, pH 7.5).
NucleoSpin RNA Clean-Up kits are also recommended for cleanup of in vitro-transcribed RNA and aminoallyl mRNA, as well as biotinylated and fluorescently-labeled RNA. Purified RNA is ready for use in applications such as enzymatic labeling reactions (including dye incorporation), RT-PCR, and DNA/RNA-based chip hybridizations.
The integrity of purified RNA can be examined by denaturing agarose gel electrophoresis.
RNA-containing samples are mixed with a solution containing large amounts of chaotropic ions. This solution immediately inactivates RNases—which are present in virtually all biological materials—and creates appropriate binding conditions which favor adsorption of RNA to the silica membrane. Simple washing steps remove salts, metabolites, organics like phenol, and cellular debris. Pure RNA can then be eluted with RNase-free water.
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