Haynes Lab:Notebook/Engineering PC-TFs/2012/10/19

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(Autocreate 2012/10/19 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
Current revision (18:57, 23 October 2012) (view source)
(Summary)
 
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==Summary==
==Summary==
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*  
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*Retrieve gel slices from fridge.
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*Add 3 volumes of ADB Buffer to each volume of gel. (Each gel slice is assumed to be 200 mg)
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*Therefore, add 600 μL of ADB Buffer.
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*Incubate at 55°C for 10 minutes.
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*Load melted agarose solution into Spin Column in a collection tube.
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*Centrifuge at max speed for 30 seconds.
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*Discard flow-through.
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*Add 200 μL of DNA wash buffer to the spin columns and centrifuge for 30 seconds.
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*Place spin column in a 1.5 mL tube. Add 10μL of DNA Elution Buffer to elute the DNA.
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*Place tubes into freezer box.
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Summary

  • Retrieve gel slices from fridge.
  • Add 3 volumes of ADB Buffer to each volume of gel. (Each gel slice is assumed to be 200 mg)
  • Therefore, add 600 μL of ADB Buffer.
  • Incubate at 55°C for 10 minutes.
  • Load melted agarose solution into Spin Column in a collection tube.
  • Centrifuge at max speed for 30 seconds.
  • Discard flow-through.
  • Add 200 μL of DNA wash buffer to the spin columns and centrifuge for 30 seconds.
  • Place spin column in a 1.5 mL tube. Add 10μL of DNA Elution Buffer to elute the DNA.
  • Place tubes into freezer box.


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