Team:Paris Saclay/Protocols/Electro elution of DNA from agarose gel
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+ | =Electro elution of DNA from agarose gel= | ||
+ | # Run samples on agarose gel as usual. When fragments are well separated, slice out bands of interest with a razor blade under the UV plate. Make slices as thin as possible (trim off excess agarose).Slices can be stored in microcentrifuge tubes in the refrigerator if necessary. | ||
+ | # Fill the vat with TAE 0.5X buffer | ||
+ | # Close one of the wells of the saline bridge with a semi-permeable membrane, put the DNA in the large well | ||
+ | # Apply a 400V current after closing the vat, for 30min | ||
+ | # Reverse the polarity for 20s, pipet the solution from the small well and put it in a clean 1.5 ml tube | ||
+ | # Add 200µL of water and 600µL of phenol/chloroform/isoamyl alcohol ratio 25/24/1 | ||
+ | # Vortex for 1 min | ||
+ | # Pipet the aqueous phase | ||
+ | # Precipitate the DNA with ethanol | ||
{{Team:Paris_Saclay/protocols_footer}} | {{Team:Paris_Saclay/protocols_footer}} |
Latest revision as of 00:02, 18 October 2014
Electro elution of DNA from agarose gel
- Run samples on agarose gel as usual. When fragments are well separated, slice out bands of interest with a razor blade under the UV plate. Make slices as thin as possible (trim off excess agarose).Slices can be stored in microcentrifuge tubes in the refrigerator if necessary.
- Fill the vat with TAE 0.5X buffer
- Close one of the wells of the saline bridge with a semi-permeable membrane, put the DNA in the large well
- Apply a 400V current after closing the vat, for 30min
- Reverse the polarity for 20s, pipet the solution from the small well and put it in a clean 1.5 ml tube
- Add 200µL of water and 600µL of phenol/chloroform/isoamyl alcohol ratio 25/24/1
- Vortex for 1 min
- Pipet the aqueous phase
- Precipitate the DNA with ethanol