Simultaneous isolation of RNA, DNA, and protein from one lysate—NucleoSpin TriPrep
The NucleoSpin TriPrep kit allows you to isolate RNA, DNA, and protein simultaneously from one lysate, allowing sensitive gene expression analysis for small, rare, or precious samples. Parallel RNA, DNA, and protein extraction from the same experimental sample is increasingly in demand by researchers interested in gene regulation mechanisms, such as knockdown of mRNA expression by siRNAs and other processes that influence protein expression.
The NucleoSpin TriPrep kit allows you to isolate RNA, DNA, and protein simultaneously from one lysate, allowing sensitive gene expression analysis for small, rare, or precious samples. Parallel RNA, DNA, and protein extraction from the same experimental sample is increasingly in demand by researchers interested in gene regulation mechanisms, such as knockdown of mRNA expression by siRNAs and other processes that influence protein expression. This method is preferable to isolating RNA, DNA, and protein from different sample aliquots, which is difficult when the amount of starting material is limited and necessitates proof of sample equality to provide reliable results.
The nucleic acids are separated by sequential elution steps. First the DNA is eluted with DNA Elute, a low-ionic strength buffer, while the RNA is still bound to the column. DNA elution is followed by wash steps, on-column DNA digestion (to remove residual DNA contamination), and elution of high-quality RNA. Proteins in the flowthrough are precipitated by a special buffer (Protein Precipitator), pelleted by centrifugation, washed, and then dissolved in Protein Solving Buffer. Isolated RNA and DNA are of high quality and ready to use for all common downstream applications. Isolated protein may be used directly for quantification, SDS-PAGE, or Western blot analysis.
Overview
- Convenient single-column isolation of RNA, DNA, and protein
- Provides high-quality RNA and DNA, ready to use for all typical downstream applications
- High protein yield is independent of protein size, localization, modification, etc.
- Complete kit includes shredders, on-column rDNAse, and a buffer for dissolving a variety of proteins
- Preserves protein primary structure and post-translational modifications (e.g., protein phosphorylation), allowing protein analysis without additional inhibitors (e.g., proteinase or phosphatase inhibitors)
More Information
| Technology | Silica membrane technology |
| Format | Mini spin columns |
| Starting material | <5 x 106 cultured cells, <30 mg human/animal tissue, <100 mg plant tissue |
|
RNA |
DNA |
Protein |
|
| Fragment size |
>200 nt |
<30 kb |
15–300 kb |
| Typical yield |
<70 µg |
<6 µg |
<1,200 µg |
| A260/280 |
1.9–2.1 |
1.7–1.9 |
— |
| Typical RIN (RNA integrity number) |
>9 |
— |
— |
| Elution volume (RNA and DNA) resolubilization volume (protein) |
40–120 µl |
100 µl |
10–100 µl |
| Preparation time |
30 min/6 preps |
45 min/6 preps (RNA + DNA) |
35 min/6 preps |
| Binding capacity |
200 µg |
10 µg* |
— |
*The binding capacity for DNA is <10 μg, and depends strongly on the amount of RNA bound to the membrane.
Applications
- Rapid purification of total RNA, DNA, and protein from small and precious samples—without splitting samples
- Gene expression profiling, analysis of transgenic organisms, drug screening, and genotyping
- Reliable interpretation of RNA, DNA, and protein amounts
- Isolation of RNA and DNA that is ready to use for all typical downstream applications
- RNA, DNA, and protein purification from a broad spectrum of starting materials, including cultured cells, tissue, bacteria, yeast, and plants
- Purification of proteins that are ready to use for SDS-PAGE/Western blotting
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Co-purify RNA, DNA, and protein from one sample
Simultaneous RNA, DNA, and protein extraction from the same experimental sample is becoming increasingly important for researchers studying gene regulation mechanisms, such as siRNA-mediated mRNA knockdown. Usually, different aliquots of sample are used to isolate nucleic acids and protein, but this is difficult when working with small, rare, or precious samples. Additionally, this type of processing necessitates proof of sample equality to ensure the data is reliable. These issues can be solved by parallel isolation of RNA, DNA, and protein from unsplit samples using the NucleoSpin TriPrep kit.
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