Difference between revisions of "Team:Freiburg/Protocols/Gel Extraction"
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Revision as of 16:08, 14 September 2015
Gel Extraction
Protokoll zur Aufreinigung von DNA nach Gel-Elektrophorese
Protocol on the purification of DNA after gel electrophoresis
Protocol by QIAGEN, modified
material: Kit (z.B. QIAGEN …)
duration: 90 min
- cut out DNA band (use UV protection) and transfer into Eppendorff tube (weight gel slice)
- 500 µL QG-Buffer (3/1000 of the mass of the gel slice)
- incubate 10 min @ 50°C, vortex from time to time
- load onto the column
- centrifuge 1 min, full speed @ RT (discard flow through)
- 500 µL QG-Buffer
- centrifuge 1 min, full speed @ RT (discard flow through)
- 750 µL PE washing buffer* - incubate 2-5 min @ RT - centrifuge 1 min, full speed @ RT - turn tubes 180° - centrifuge 1 min full speed @ RT - transfer column into Eppendorff tube - pipet 25 µL water on the membran (may be warmed up before) - incubate 10 min RT - incubate 3 min, 55°C - centrifuge** 1min, full speed RT
⇒ DNA is transferred into the Eppendorff tube
To increase yield two bands at once may be loaded onto the column
