Team:Freiburg/Protocols/Gel Extraction

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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

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1. cut out DNA band (use UV protection) and transfer into Eppendorff tube (weight gel slice)
2. 500 µL QG-Buffer (3/1000 of the mass of the gel slice)
3. incubate 10 min @ 50°C, vortex from time to time
4. load onto the column
5. centrifuge 1 min, full speed @ RT (discard flow through)
6. 500 µL QG-Buffer
7. centrifuge 1 min, full speed @ RT (discard flow through)
8. 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

protocols old