This kit provides for rapid spin column isolation and purification of total RNA and genomic DNA simultaneously from a single plant sample without splitting the lysate. Norgen’s plant lysis solution is highly robust and effective over a wide range of plants including challenging samples. The total RNA and genomic DNA are both column purified in under 30 minutes. Since RNA and DNA are isolated without splitting the lysate, variability and inconsistent results are reduced. All sizes of RNA including microRNA are recovered without the need for phenol. Optional on-column DNase and RNase treatments provide flexibility to isolate DNA-free RNA or RNA-free DNA respectively. Isolated nucleic acids are of a high quality and yield, and are ready for downstream use including PCR, qPCR, RT-PCR, qRT-PCR, sequencing and more.
Background
It is often necessary to isolate total RNA and genomic DNA from a single plant sample, such as for studies of gene expression, mutant or transgenic plant characterization, and host plant-pathogen characterization. This is of great benefit when isolating RNA and DNA from precious, difficult to obtain or very small samples. Furthermore, gene expression analysis will be more reliable since the RNA and DNA are derived from the same sample, therefore eliminating inconsistent results.
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| Kit Specifications | |
| Column Binding Capacity | 50 μg for RNA 15 μg for genomic DNA |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Maximum Amount of Starting Material: Plant Tissues Plant Cells | 100 mg 5 x 106 |
| Time to Complete 10 Purifications | 30 minutes |
| Average Yields* Peach Leaves (100 mg) | 40 μg RNA, 5 μg gDNA |
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 2 years in their unopened containers.
| Component | Cat. 24400 (50 preps) |
|---|---|
| Lysis Buffer M | 40 mL |
| Binding Buffer I | 7 mL |
| Solution WN | 18 mL |
| Wash Solution A | 38 mL |
| Elution Buffer E | 20 mL |
| Enzyme Incubation Buffer B | 6 mL |
| Filter Columns | 50 |
| Spin Columns | 50 |
| Collection Tubes | 100 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
This product provides a fast and easy way to purify DNA from plant and fungal tissue. Up to 100 mg of tissue can be processed. Easy-to-use Plant procedures provide pure total DNA (genomic,mitochondrial, and chloroplast) for reliable PCR and southern blotting in less than 1 hour. Purification requires no phenol or chloroform extraction oralcohol precipitation and involves minimal handling.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from100 mg simple plant |
| Applications | PCR, SSR, AFLP, RAPD and southern blot, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Economic plant samples |
| Sample amount | Fresh / frozen plant samples: ≤100mggDried plant / seed samples: ≤20mg |
| Elution volume | ≥30μl |
| Time per run | 30-50 minutes |
| Liquid carrying volume per column | 800μl |
| Binding yield of column | 100μg |
Plantmaterial is first mechanically disrupted and then lysed by addition of lysis buffer and incubation. RNase A in the lysis buffer digests the RNA in the sample. After lysis, proteins and polysaccharides are salt-precipitated.Binding buffer and ethanol are added to the cleared lysate to promote binding of the DNA to the HiPure membrane. The sample is then applied to a column and then centrifuged. DNA binds to the membrane, while contaminants such as proteins and polysaccharides are efficiently removed by 2 wash steps. Pure DNA is eluted in a small volume of low-salt buffer or water.
Kit Contents
| Contents | D316402 | D316403 |
| Purification Times | 50 Preps | 250 Preps |
| RNase A | 10 mg | 50 mg |
| Protease Dissolve Buffer | 1.8 ml | 5 ml |
| Buffer SPL | 30 ml | 150 ml |
| Buffer PS | 10 ml | 50 ml |
| Buffer GW1 | 22 ml | 110 ml |
| Buffer GW2* | 12 ml | 2 x 50 ml |
| Buffer AE | 15 ml | 60 ml |
| HiPure gDNA Mini Columns | 50 | 2 x 125 |
| 2 ml Collection Tubes | 50 | 2 x 125 |
Storage and Stability
RNase A should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Experiment Data
| Name | CAT NO | Fresh / frozen tissue amount | Dried tissue amount | Column type | Elution volume | Yield (muscle) |
| HiPure Plant DNA Mini Kit | D3187 | 150mg | 40mg | 1.5ml column | 50 – 100μl | 3-70μg |
| HiPure HP Plant DNA Maxi Kit | D3163 | 5g | 1g | 50ml column | 0.7 – 3ml | 75-1250μg |
| HiPure SF Plant DNA Kit | D3164 | 100mg | 20mg | 1.5ml column | 50 – 100μl | 3-50μg |
| HiPure SF Plant DNA 96 Kit | D3167 | 50mg | 15mg | 96 well plate | 75 – 150μl | 3-30μg |
This product provides a fast and easy way to purify DNA from plant and fungal tissue. Up to 100 mg of tissue can be processed. Easy-to-use Plant procedures provide pure total DNA (genomic,mitochondrial, and chloroplast) for reliable PCR and southern blotting in less than 1 hour. Purification requires no phenol or chloroform extraction oralcohol precipitation and involves minimal handling.
