Agarose Gel Extraction Kit
฿4,700.00 – ฿10,700.00
For general laboratory use.
Shipping: shipped at ambient temperature
Storage Conditions: store at ambient temperature
Shelf Life: 12 months
Description:
Agarose Gel Extraction Kit is designed for high-yield recovery of DNA from agarose gel with simultaneous removal of primer-dimers, 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 kb. It requires no organic extractions or precipitation and guarantees high and stable recovery rates.
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
To be provided by you:
96-99 % Ethanol
Isopropanol
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.
Buffer | PP-202S 50 preps |
PP-202L 250 preps |
Extraction Buffer | 75 ml | 2x 185 ml |
Activation Buffer | 6 ml | 30 ml |
Washing Buffer | add 64 ml Ethanol (final volume 80 ml) |
add 160 ml Ethanol to each bottle (final volume 200 ml each) |
Elution Buffer | 5 ml | 25 ml |
The additional use of Isopropanol enhances yield and 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.
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.
- Add 1 volume Isopropanol per gel volume to the dissolved gel and mix well.
- For purification of DNA fragment sizes smaller than 200 bp or larger than 5 kbp increase the amount of Isopropanol to 2 volumes.
3 Column Activation:
- Place a Spin Column into a 2 ml collection tube.
- Add 100 μl of Activation Buffer into the Spin Column.
- Centrifuge at 10,000 g for 30 sec in a micro-centrifuge.
4 Column Loading:
- Apply the sample mixture from step 2 into the activated Spin Column.
- Centrifuge at 10,000 g for 30 sec in a micro-centrifuge.
- Discard the flow-through.
5 Column Washing:
- Place the DNA loaded Spin Colum into the used 2 ml tube.
- Apply 700 μl of Washing Buffer 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 and 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.
Vendor | Jena Bioscience |
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