| Technology | Silica membrane | |
| Starting material | <5 x 106 cultured cells <30 mg human/animal tissue <100 mg plant tissue |
|
| Typical yield | DNA RNA Protein |
<6 µg <70 µg <1,200 µg |
| Typical A260/280 | DNA RNA |
1.7–1.9 1.9–2.1 |
| Preparation time | RNA/DNA/protein RNA/DNA Protein |
~60–75 min ~45 min ~35 min |
Tech Note
NucleoSpin TriPrep: parallel isolation of RNA, DNA, and protein from one sample
- Convenient one-column preparation of RNA, DNA, and protein from diverse sample types
- High-quality RNA and DNA, ready to use for downstream applications
- High protein yield independent of protein size, localization, modification, etc.
- Complete kit includes NucleoSpin Filters (shredders) for optimal lysis, rDNase for on-column DNA digestion, and Protein Solving Buffer for dissolving all types of proteins
NucleoSpin TriPrep allows you to isolate DNA, RNA, and protein without splitting the sample prior to extraction. The fractions are obtained from the exact same sample, and not from three similar portions of one sample. This is especially valuable for unique, small, and precious samples.
Product summary
Diverse sample types
RNA/DNA/protein extraction from a wide variety of starting materials
RNA, DNA, and protein were isolated in parallel from the same source. The range of starting materials includes tissues, cells, plant materials, yeast, and bacteria.
| Lane | Starting material | Sample amount |
| 2 | Human cells (HeLa) | 106 cells |
| 3 | Mouse liver | 3 mg |
| 4 | Fish (Zebrafish) | 30 mg |
| 5 | Plant root (garden cress) | 100 mg |
| 6 | Plant leaves (garden cress) | 100 mg |
| 7 | Yeast | 108 cells |
| 8 | Bacteria | 109 cells |
Total DNA
Figure 1. Total DNA fractions, 1% TAE agarose gel electrophoresis*. High-quality DNA was eluted in 100 µl DNA Elute following the NucleoSpin TriPrep procedure. The DNA is of high molecular weight and purity. A260/A280 ratios are in the range of 1.81–1.94.
Total RNA
The isolated RNA is of high integrity. RIN (RNA integrity number) was measured for mammalian samples. RIN of RNA isolated from HeLa cells was 9.5, RIN of RNA from mouse liver sample was 9.1.
Figure 2. Total RNA fractions analyzed on an Agilent 2100 Bioanalyzer/RNA 6000 Nano Kit*. The isolated RNA is of high integrity. RIN (RNA integrity number) was measured for mammalian samples. RIN of RNA isolated from HeLa cells was 9.5, RIN of RNA from mouse liver sample was 9.1.
Total Protein
Figure 3. Total protein fractions, SDS-PAGE analysis*. Isolated protein is dissolved in Protein Solving Buffer, provided with the kit. The protein is ready-to-use for SDS-PAGE analysis, as well as for Western blotting.
* Figures are compiled from different gels and runs, aligned to the corresponding length markers.
Protocol schematic
Samples are lysed by incubation in a buffer containing chaotropic salts. This will not only destroy the cell structure of the sample but simultaneously inactivate enzymes such as RNases, phosphatases, and proteases, and prevent degradation of nucleic acids and proteins. At the same time, the buffer creates appropriate binding conditions for RNA and DNA.
Nucleic acids and protein from the sample can be separated by simply centrifuging the lysate through the NucleoSpin TriPrep spin column: RNA and DNA are bound to the silica membrane while the protein is recovered in the column flowthrough.
The nucleic acids are separated by sequential elution steps. First the DNA is eluted with DNA Elute, a low ionic strength buffer, while RNA is still bound to the column. RNA is isolated, following wash steps, on-column DNA digestion (to remove residual DNA), and elution.
Proteins in the flowthrough are precipitated by a special buffer (Protein Precipitator), pelleted by centrifugation, washed, and dissolved in Protein Solving Buffer.
Isolated RNA and DNA are of high quality and directly suitable for all common downstream applications. Isolated protein is immediately suitable for quantification, SDS-PAGE, and Western blot analysis.
Figure 4. Outline of the NucleoSpin TriPrep workflow.
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