Corum:Gel Purification

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Contents

Overview

Gel extraction with Invitrogen Purelink Gel Extraction Kit.

Materials

  • DNA embedded in gel
  • Purelink column
  • Gel solubilization buffer
  • Isopropynol
  • Wash buffer
  • Elution buffer or sterile ddH2O.

Procedure

  1. Excise DNA from gel with razor blade. Place in 1.5 mL tube and weigh gel piece. Let V be the weight of the piece in mg.
  2. Add 3V μL Gel Solubilization Buffer. Incubate 55 °C 10 min or until gel is completely dissolved.
  3. After gel is completely dissolved, incubate 55 °C additional 5 min.
  4. Incubate RT 2 min.
  5. Add 1V μL isopropynol. Mix and spin.
  6. Apply to column. Centrifuge max speed 1 min. Discard flowthrough. (Maximum application volume is 700 μL, so for larger volumes, repeat until all of the DNA is bound to column membrane.)
  7. Apply 700 μL wash buffer to column. Centrifuge max speed 1 min. Discard flowthrough.
  8. To dry, centrifuge max speed 3 min. Place column in a new 1.5 mL tube.
  9. Apply 50-100 μL elution buffer or sterile ddH2O to column membrane. Avoid touching membrane with tip. Incubate RT 10 min.
  10. To elute, centrifuge 1 min max speed. Discard column. DNA is now in bottom of 1.5 ML tube.
  11. Quantify DNA sample by quantifluore.
  12. Label side of tube with the following:
    • [DNA] in nM
    • "gel purified"
    • date
  13. Store at -20 °C.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

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References

Invitrogen Purelink Gel Extraction Kit manual.

Contact

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Digital Signature

  • SC 17:57, 25 July 2012 (EDT):
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