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Agarose Gel Extraction Kit | DNA Cleanup Cellco
Agarose Gel Extraction Kit
Ficha técnica
| Código | Agarose Gel Extraction Kit |
| Categoria | DNA Cleanup |
| Marca | Cellco |
Descrição Geral
Agarose Gel Extraction Kit | DNA Cleanup - Cellco
Mix de Exonuclease/Fosfatase Alcalina para clean-up de produtos de PCR
For in vitro use only!
Envio:
Shipped at ambient temperature
Condições de armazenamento:
Store at ambient temperature
Validade:
12 months
Descrição:
Agarose Gel Extraction Kit is designed for high-yield recovery of DNA from agarose gel with simultaneous removal of primerdimers, primers, nucleotides, proteins, salt, agarose, ethidium bromide, and other impurities. The preparation is based on a silica-membrane technology for binding DNA in high-salt and elution in low-salt buffer. The kit provides a simple and efficient way to purify DNA in a size range between 100 bp and 10 kbp. It requires no organic extractions or precipitation and guarantees high and stable recovery rates.
Kit Content:
- Extraction Buffer
- Activation Buffer
- Washing Buffer (before use, add 96-99 % Ethanol as indicated on the bottle)
- Elution Buffer
- Spin Columns
- 2 ml Collection Tubes
Additional Materials Required:
- 96-99% Ethanol
- Isopropanol (optional)
- 1.5 ml microtubes
Preparation Procedure:
The agarose gel is dissolved in the chaotropic Extraction Buffer followed by a simple binding, washing, and eluting procedure. Before start, add 96-99 % Ethanol to the Washing Buffer as indicated on the bottle.
The additional use of Isopropanol is recommended for fragments smaller than 200 bp or larger than 5 kbp. The optional secondary washing step minimizes the salt content of the purification product but may significantly reduce the yield of DNA fragments <200 bp.
| Buffer | DPK-105XS 10 preps | DPK-105S 50 preps | DPK-105L 250 preps |
| Extraction Buffer | 15 ml | 75 ml | 2 x 185 ml |
| Activation Buffer | 1.2 ml | 6 ml | 30 ml |
| Washing Buffer | add 12 ml Ethanol (final volume 15 mL) | add 64 ml Ethanol (final volume 80 mL) | add 160 ml Ethanol to each bottle (final volume 200 mL) |
| Elution Buffer | 1 ml | 5 ml | 25 ml |
1. Excision of the Gel:
• Cut the area of gel containing the DNA fragment.
• Transfer the excised gel to a clean 1.5 ml microtube.
2. Sample Preparation:
• Add 3 volumes of Extraction Buffer to 1 volume of the sliced gel.
For example, add 300 µl Extraction Buffer to each 100 mg (approx.100 µl) gel. For gels containing >2.5 % agarose, add 6 volumes of Extraction Buffer per gel volume.
• Incubate at 60 °C for 10 min with occasional mixing to ensure gel dissolution.
• For DNA fragment sizes smaller than 200 bp or larger than 5 kbp and to enhance yield add 1 volume Isopropanol per gel volume to the dissolved gel and mix well.
3. Column Activation:
• Place a Spin Column into a 2 ml collection tube.
• Add 100 µl of Activation Buffer into the Binding Column.
• Centrifuge at 10,000 g for 30 sec in a micro-centrifuge.
4. Column Loading:
• Apply the sample mixture from step 2 into activated Spin Column.
• Centrifuge at 10,000 g for 30 sec.
• Discard the flow-through.
4. Column Washing:
• Place the DNA loaded Spin Column into the used 2 ml tube.
• Apply 700 µl of Washing Buffer (containing Ethanol) to the Spin Column.
• Centrifuge at 10,000 g for 30 sec and discard the flow-through.
Optional Secondary Washing:
Recommended only for DNA >200 bp, if highly purified DNA (for DNA sequencing, transfection etc.) is required.
• Add 700 µl of Washing Buffer to the Spin Column.
• Centrifuge at 10,000 g for 30 sec and discard the flow-through.
• Centrifuge again for 2 min to remove residual Washing Buffer.
6. Elution
• Place the Spin Column into a clean 1.5 ml microtube (not provided in the kit).
• Add 30-50 µl Elution Buffer or dd-water to the center of the column membrane.
• Incubate for 1 min at room temperature.
• Centrifuge at 10,000 g for 1 min to elute DNA.
Informações Detalhadas
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