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Si-Mag™ AG1 Kit


Agarose Gel DNA Extraction and Purification Magnetic Beads Kit
This kit provides a simple, rapid and efficient method for the recovery and purification of DNA directly from Agarose gel (100 bp to 50 kb) with typical recovery efficiency up to 85%. The resulting product can be directly used for sequencing, restriction digestion, or PCR and other downstream experiments. In addition, the kit can be used to concentrate DNA.
The kit will work with a 96 well round bottom plates if a special magnetic frame is used. The kit
can also be used with a variety of automatic nucleic acid extraction instruments and workstation.

Catalog #

Si-Mag-AG1

Precautions

  • Wear protective gloves, clothing, eye, and face protection. Wash hands thoroughly after handling.
  • Avoid freeze/thaw cycles and centrifugation which could damage the beads.
  • Vortex beads for about 10 seconds and mix them well with DNA containing material to ensure best performance.
  • Vortex samples for about 10 seconds before adding.
  • Elute DNA from the beads completely.

Kit Components included

  1. Si-Mag magnetic beads 5.5 mL;
  2. Gel solvent buffer 34 mL (for purification of DNA from Agarose gel);
  3. Wash Solution 38 mL (add 25 mL of Isopropanol before use);
  4. Elution Buffer 4 mL.

Materials needed but not provided with the kit

  • 80% Ethanol in water.
  • Si-Mag Magnet (sold separately) or other magnetic racks compatible with vials used.
  • Isopropanol (ACS grade).
  • RNAse 100 mg/mL solution to remove RNA.

Storage

Magnetic beads should be stored at 2-8°C but other kit reagents need to be stored at room temperature. Lysis solution may turn cloudy if store in th cold room and to clear it up place the bottle into a water bath at 37°C.

Protocol

  1. Sample preparation. Add 500 uL of gel solvent buffer and a gel slice of up to 400 mg into a clean Eppendorf tube. For a gel slice larger than 400 mg, add at least 800 ul of gel solvent buffer. Incubate at 65°C for 10 min or until the gel is completely dissolved. Vortex the tube periodically to ensure complete dissolving.

  2. Transfer all content to an Eppendorf tube and then add 50 uL of magnetic beads, mix well and incubate 3-5 min at RT. Put Eppendorf tube onto the Si-Mag magnet rack for 20 seconds.
    NOTE: The estimated recovery of DNA is about 2 ug per 50 uL of beads.
  3. Remove supernatant by holding the magnet rack upside down or by pipetting.
  4. Wash the beads with 500 uL of Wash Solution. Make sure the beads get completely resuspended by vortexing and then repeat step 3.
  5. Wash the beads with 500 uL of 80% ethanol.
  6. Make sure the beads get completely resuspended by vortexing and then repeat step 3.
  7. Dry the beads at 55°C for 8 min leaving the tube open. Do not over-dry the beads.
  8. Elute the DNA from beads with 35 uL of elution buffer, incubate for at least 2 min and then vortex at full speed for 1 min. Alternatively, incubation at 60°C for 2 min may improve the recovery for DNA larger than 3 kb.
  9. Remove beads by using magnet rack, pipette DNA out and transfer to a clean tube.
  10. Store purified DNA at -20°C for long-term storage.


 

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