PCR purification: Difference between revisions
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{{Protokoll-Mix|Inhalt= | {{Protokoll-Mix|Inhalt=“QIAquick gel extraction/PCR purification” – Kit with the PB buffer | ||
1.5 mL labeled Eppendorf tubes|titel=Materials}} | |||
== Procedure == | |||
* The purification of PCR fragments is obtained with the aid of a kit | |||
* Add 5 volumes of PB buffer to the PCR product and transfer the whole volume to a column | |||
* Centrifugation: 60 seconds, 13000 rpm | |||
* Discard the flowthrough and add 0.75 mL buffer PE to the column and centrifuge again | |||
* Discard the flowthrough and centrifuge again for 60 sec | |||
* The column is added to a sterile, labeled Eppendorf tube. | |||
* Add 30 μl EB-buffer directly to the column, incubate 1 min and elute by centrifugation (60 sec, 13000 rpm). | |||
* Store the eluate on ice | |||
Latest revision as of 11:22, 30 May 2025
Materials
“QIAquick gel extraction/PCR purification” – Kit with the PB buffer
1.5 mL labeled Eppendorf tubes
Procedure
- The purification of PCR fragments is obtained with the aid of a kit
- Add 5 volumes of PB buffer to the PCR product and transfer the whole volume to a column
- Centrifugation: 60 seconds, 13000 rpm
- Discard the flowthrough and add 0.75 mL buffer PE to the column and centrifuge again
- Discard the flowthrough and centrifuge again for 60 sec
- The column is added to a sterile, labeled Eppendorf tube.
- Add 30 μl EB-buffer directly to the column, incubate 1 min and elute by centrifugation (60 sec, 13000 rpm).
- Store the eluate on ice