Gel DNA Recovery
ZymoClean DNA Recovery Kit
Isolates DNA fragment from agarose gel
EXTRACTION
1 Use a sharp scalpel to remove the
desired DNA band from the agarose
gel. Place it into a 1.5 mL
microcentrifuge tube and weigh it. This
measurement is crucial for calculating
the amount of agarose dissolving
buffer (ADB) to add.
DISSOLVING
2 Use a 3:1 ratio of ADB to gel weight (e.g., 100 mg of gel
needs 300 µL of ADB). ADB contains chaotropic salts,
which denature the agarose, facilitating the release of
DNA. Incubate the tube at 55ºC for 10 minutes to
dissolve the agarose. Vortex the sample briefly to mix.
BINDING
3 Place the zymospin column into a collection
tube and transfer the melted agarose solution
to the column. The DNA will bind to the silica
membrane of the column whilst the ADB and
agarose will pass through. Allow the column to
stand for 1 minute before centrifuging at
15,000 xg for 1 minute. This drives DNA
through the membrane, where it binds. Discard
the flow through.
4 WASHING
Add 200 µL of DNA wash buffer to the
column and centrifuge for 30 seconds.
ZymoClean DNA Recovery Kit
Isolates DNA fragment from agarose gel
EXTRACTION
1 Use a sharp scalpel to remove the
desired DNA band from the agarose
gel. Place it into a 1.5 mL
microcentrifuge tube and weigh it. This
measurement is crucial for calculating
the amount of agarose dissolving
buffer (ADB) to add.
DISSOLVING
2 Use a 3:1 ratio of ADB to gel weight (e.g., 100 mg of gel
needs 300 µL of ADB). ADB contains chaotropic salts,
which denature the agarose, facilitating the release of
DNA. Incubate the tube at 55ºC for 10 minutes to
dissolve the agarose. Vortex the sample briefly to mix.
BINDING
3 Place the zymospin column into a collection
tube and transfer the melted agarose solution
to the column. The DNA will bind to the silica
membrane of the column whilst the ADB and
agarose will pass through. Allow the column to
stand for 1 minute before centrifuging at
15,000 xg for 1 minute. This drives DNA
through the membrane, where it binds. Discard
the flow through.
4 WASHING
Add 200 µL of DNA wash buffer to the
column and centrifuge for 30 seconds.
