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DNA Cleanup Maxi Kit (DFM010, DFM025)
GenepHlow™ DNA Cleanup Maxi Kits were designed to recover or concentrate DNA fragments from large volume agarose gel, restriction enzyme digestion or PCR reaction products. Gel/PCR Buffer (yellow color indicating optimal pH≤7.5) is premixed with a pH indicator to ensure optimal pH, facilitate DNA binding and allow for easy observation of undissolved agarose gel. If pH exceeds the optimal level (>7.5), the color of the solution will appear purple instead of yellow. 3M Sodium Acetate (pH5.0) which is included with the kit, can then be added to the solution to adjust pH and return the color to yellow. Chaotropic salt is used to dissolve agarose gel and denature enzymes while DNA fragments are bound by the glass fiber matrix of the spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified DNA fragments are eluted by a low salt Elution Buffer, TE or water. The pH indicator, salts, enzymes and unincorporated nucleotides can be effectively removed from the reaction mixture without phenol extraction or alcohol precipitation and the purified DNA is ready for use in subsequent reactions.
Quick Protocol
Advantages (Cat. # DFM010, DFM025)
- High Efficiency: up to 80% recovery
- Convenient: Gel/PCR binding buffer is premixed with a pH indicator to allow for easy determination of optimal pH
- Includes 3M Sodium Acetate (pH5.0) to adjust pH if it becomes too high
- Sample Size: up to 5 g of agarose gel slice or up to 2 ml of restriction enzyme digestion product or PCR product
- Broad Fragment Size Range: 100 bp–10 kb
- Operation Time: 24 minutes for enzymatic reaction cleanup and 36 minutes for gel extraction
- Spin Columns: glass fiber membrane
- Elution Volume: 500-1,000 µl (50 µl dead volume)
- Binding Capacity: 200 µg
- Storage: dry at room temperature (15-25ºC) for up to 18 months without showing any reduction in performance
Applications
Fluorescent and Radioactive Sequencing, PCR, Restriction Enzyme Digestion, DNA Labeling and Ligation, Microarray, Transfection
Components
- Gel/PCR Buffer (premixed with a pH indicator)
- 3M Sodium Acetate (pH5.0)
- W1 Buffer
- Wash Bufffer
- Elution Buffer (10 mM Tris-HCl, pH8.5 at 25ºC)
- DM Columns
Quality Control
The quality of the GenepHlow™ DNA Cleanup Maxi Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. DNA fragments are purified from either agarose gel, restriction enzyme digestion products, PCR products or other aqueous solutions analyzed by electrophoresis.
GenepHlow™ DNA Cleanup Maxi Kit Functional Test Data
 | Figure 1. 200 µg of pbluescript plasmid DNA was digested by Hind III endonuclease at 37ºC for 2 hours. Following digestion, the linear DNA fragment was purified using the GenepHlow™ DNA Cleanup Maxi Kit. The unpurified DNA fragment (lane 1) and 3 replications of 5 µl purified linear DNA fragments (lanes 2-4) were loaded on 0.8% agarose gel. M = Geneaid 1 Kb DNA ladder
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| Figure 2. A pH indicator is premixed with the binding buffer to ensure optimal pH, facilitate DNA binding and allow for easy observation of undissolved agarose gel. If pH exceeds the optimal level (>7.5), the color of the solution will appear purple instead of yellow. 3M Sodium Acetate (pH5.0), which is included with the kit, can then be added to the solution to adjust pH and return the color to yellow. | ||||||||||||||||||||||||||||||||
Gel Extraction and PCR Cleanup Kits Specifications
Specifications | |||||||
| Cat. # | DFG100 DFG300 | DFC100 DFC300 | DFH100 DFH300 | DFM010 DFM025 | 96DFG04 96DFG10 | DF101 DF301 | DFL100 DFL300 |
Format | Spin Column | Spin Column | Spin Column | Maxi Spin Column | 96 Well Plate | Spin Column | Silica Matrix |
| Binding Capacity | 10 µg | 10 µg | 10 µg | 200 µg | 10 µg | 10 µg | 5 µg |
| Sample | Agarose Gel Slice | PCR Product | Agarose Gel Slice/PCR Product | Agarose Gel Slice/Restriction Enzyme Digest Product | Agarose Gel Slice | Agarose Gel Slice/PCR Product | Agarose Gel Slice/PCR Product |
| Fragment Size | 70 bp-20 kb | 70 bp-20 kb | 70 bp-20 kb | 100 bp-10 kb | 70 bp-20 kb | 40-200 bp | 100 bp-50 kb |
| pH Indicator | Yes | Yes | Yes | Yes | N/A | N/A | Yes |
| Recovery | Up to 90% | Up to 95% | Up to 95% | Up to 80% | Up to 80% | Up to 95% | Up to 95% (<10 kb) Up to 80% (10-50 kb) |
| Elution Volume | 20-50 µl | 20-50 µl | 20-50 µl | 500-1000 µl | 60-80 µl | 20-50 µl | 10-20 µl |
| Operation Time | 20 minutes | 10 minutes | <20 minutes | <25 minutes | 45 minutes | <20 minutes | <20 minutes |
Related Gel Extraction and PCR Cleanup Products
Product Name | RXNS | Catalogue Number |
100, 300 | DFG100/300 | |
100, 300 | DFC100/300 | |
100, 300 | DFH100/300 | |
10, 25 | DFM010/25 | |
4 x 96, 10 x 96 | 96DFH04/10 | |
100, 300 | DF101/301 | |
100, 300 | DFL100/300 | |
100, 300 | DF100/300 | |
| Presto™ 96 Well Gel Extraction Kit | 4 x 96, 10 x 96 | 96DFG04/10 |
| GenepHlow™ DNA Cleanup Midi Kit | 100, 300 | DFI100/300 |