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Agarose Gel Extraction Kit

Spin-column based DNA cleanup from agarose gels

Product Cat. No. Amount Price (EUR) Buy / Note
Agarose Gel Extraction Kit PP-202XS 10 preparations 12,00 Add to Basket/Quote Add to Notepad
Agarose Gel Extraction Kit PP-202S 50 preparations 45,00 Add to Basket/Quote Add to Notepad
Agarose Gel Extraction Kit PP-202L 250 preparations 180,00 Add to Basket/Quote Add to Notepad

For in vitro use only!

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.

Kit contents:
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 (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.

BufferPP-202XS
10 preps
PP-202S
50 preps
PP-202L
250 preps
Extraction Buffer15 ml75 ml2x 185 ml
Activation Buffer1.2 ml6 ml30 ml
Washing Bufferadd 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 each)
Elution Buffer1 ml5 ml25 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.
  • 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 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.

Availability Restriction: Exclusively distributed in Japan by Greiner BioOne