Tech Note
Rapid purification of high-quality RNA with low genomic DNA contamination
The NucleoSpin RNA Plus kit is designed to purify high-quality RNA from a variety of cell and tissue types. The kit integrates an innovative feature, the NucleoSpin gDNA Removal Column, which rapidly and efficiently removes genomic DNA without the need for DNase.
- NucleoSpin gDNA Removal Column—no time consuming DNase digestion steps
- High-quality RNA—lysis buffer results in immediate inactivation of RNases
- Convenient handling—lysate clearing and gDNA removal with one column in one step
- New efficient lysis buffer—no beta-mercapthoethanol or TCEP necessary
Product summary
Protocol schematic
One of the most important critical aspects of any RNA isolation protocol is to prevent degradation of the RNA. When using NucleoSpin RNA Plus cells and tissues are first lysed by incubation in a chaotropic ion lysis buffer solution, which immediately inactivates RNases. The lysate is added to the NucleoSpin gDNA Removal Column (yellow rings) to clarify the lysate and to remove contaminating gDNA without the need for a DNase I step. After the addition of the Binding Solution to the flow-through, the RNA is bound to the NucleoSpin RNA Plus Column (light blue rings). Two subsequent wash steps remove salts, metabolites, and macromolecular cellular components. High-quality RNA is eluted with RNase-free H2O.
High yield
RNA was isolated from HeLa cells (5 x 106) and mouse kidney (20 mg) with the NucleoSpin RNA Plus (blue) and competitor Q's kit (red). The RNA recovery was quantified by qRT-PCR. The CT values are same or lower for MN, indicating similar or better yield for the MN kit.
Genomic DNA removal
NucleoSpin RNA Plus (blue) and competitor Q's kit (red) were used for the isolation of RNA from HeLa cells (5 x 106) and mouse kidney (20 mg). Residual genomic DNA was measured by qPCR. The MN kit shows higher CT values indicating more efficient removal of genomic DNA.
High RNA quality (RIN)
The RNA integrity number (RIN) provides a numerical assessment of the quality of RNA samples, it is based on a scale from 1 to 10, with 1 being the most degraded and 10 being the most intact. To compare the quatity of RNA produced using NucleoSpin RNA Plus (blue) and Competitor Q's kit (red) RNA from mouse kidney tissue (10 mg) was isolated and amounts were analyzed using an Agilent Bioanalyzer to determine the RIN. The RIN value from RNA purified using NucleoSpin RNA Plus was far higher, indicating better quality RNA.
Ordering information
Universal reagent for RNA isolation
NucleoZOL is a highly efficient RNA isolation reagent for extracting high-quality miRNA and total RNA from a variety of starting materials, including cells, tissues, and liquids of human or animal origin, as well as plants, yeast, bacteria, viruses, and other sources.
See the data RNA purification kitsTakara Bio USA, Inc.
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