Zymoclean Gel DNA Recovery Ki

Highlights

• Ultra-pure DNA recovered in 6 ul of water ensures that your ligations, sequencing, etc. work every time.
  
• New buffer system works with either TAE or TBE buffers. No need to adjust pH as in other kits.
  
• 75 bp to 23 Kb DNA size range. No DNA shearing as found in glass milk or cation rasin milk procedure.
  
• Very simple procedure: Dissolve gel. Spin through Zymo-Spin column. Wash and elute. Whole procedure takes less than 15 minutes.
  
• 70-95% recovery rate.

General Description
Zymoclean Gel DNA Recovery Kit is based on our newly formulated chemistry and Zymo-Spin columns for simple, efficient and ultra-pure DNA recovery from agarose gels. This reliable method easily recovers DNA from agarose gels into small volumes, 6-8 ul, of pure water for convenient downstream reactions. The chemistry and the column were developed and tested for recovering DNA to give the most efficient ligation and sequencing reading. The recovered DNA was also tested for other procedures such as probe labeling, kinase reactions, PCR amplification, etc. The new single buffer system eliminates the need to add additional isopropanol or pH adjustments. Applicable DNA sizes range from 75 bp to 23 Kb. There is no DNA shearing or inhibitor carry-over. The DNA recovery rate is 70-95%.

Protocol
Before starting: Add 16 ml (64 ml for D4002 of 200 gel DNA recovery kit) of 100% ethanol to the Wash Buffer Concentrate to make final Wash Buffer.

1. Excise the DNA fragment from the agarose gel with a razor blade or a scalpel and transfer to a 1.5 ml tube.
Gel volume excised should be as small as possible.
2. Add 3 volumes of ADB-Buffer* (Agarose Dissolving Buffer) to each volume of agarose gel.
Example: For 100 ul (or mg) of agarose gel, add 300 ul of ADB buffer.
For DNA fragments >8 Kb, add one additional volume of water for better recovery. Example: 100 ul agarose, 300 ul of ADB Buffer and 100 ul of water.
3. Incubate at 55°C for 5-10 minutes until the gel is completely dissolved.
Do no incubate above 60°C. Higher temperatures may result in no DNA recovery. Incubation time depends on the size of the gel. It is important that the gel is dissolved completely. Mixing during the incubation period aids the gel dissolving process.
4. Load the melted agarose solution into a Zymo-Spin Column and place column into a 2 ml collection tube.
The capacity of the column is 600 ul. Load twice if the volume is more than 600 ul. The capacity of the collection tube is 600 ul with the column inserted. Empty collection tube whenever necessary if the volume is more than 600 ul to prevent the contamination of column by flow-through.
5. Centrifuge at full speed (>10,000 g) for 5-10 seconds. Discard the flow-through.
6. Add 200 µl of Wash Buffer to the column. Spin at top speed for 5-10 seconds. Add another 200 ul of Wash Buffer. Spin at top speed for 30 seconds.
Important! A longer spinning duration for this last wash is necessary for complete removal of wash buffer residues.
7. Add 6-8 ul of water directly to the column matrix. Place column into a 1.5 ml tube. Spin briefly to elute the DNA

Note: TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) can also be used for the elution step instead of water. Waiting for one minute after adding water or TE before centrifuging can improve DNA recovery rate for larger sizes of DNA (>6 Kb).

DNA is now ready for your experiments.

*Contains Chaotropic reagents. Irritant. Please use proper safety precautions with this reagent.

Zymo Research, 625 West Katella Ave, Suite 32, Orange, CA 92867-4619. Tel: 714-288-9682; Fax: 714-288-9643.