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DNA extraction and precipitation with ethanol / isopropanol: Difference between revisions
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DNA extraction and precipitation with ethanol / isopropanol (view source)
Revision as of 11:48, 28 October 2012
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== Sample collection == | |||
* Collect a small amount of cells by gently scraping the interior of your cheek. | |||
* Transfer the collected cells to a 1.5 ml tube. | |||
== Novel experimental procedure for DNA extraction (28/10/12) == | |||
(This has not yet been tested) | |||
* Add to the cell sample 200 µl of Lysis Buffer (10 mM Tris-HC1 (pH 8.0), 1 mM EDTA, 1% Triton X-100). | |||
* Incubate solution at 94˚C for at least half an hour (better 1h). | |||
* Spin in microcentrifuge at 13,000 rpm for 10 min. | |||
* Collect supernatant in a novel tube and discard pellet. Procede with proteinase K treatment and ethanol precipitation. | |||
== Proteinase K treatment and ethanol precipitation == | |||
* Get DNA sample from previous extraction. | * Get DNA sample from previous extraction. | ||
* Add 1 part proteinase K (1mg/ml stock solution) to 3 parts sample. Incubate at 50˚C for 1 hour and then heat-inactivate proteinase K at 94˚C for 10 min. | * Add 1 part proteinase K (1mg/ml stock solution) to 3 parts sample. Incubate at 50˚C for 1 hour and then heat-inactivate proteinase K at 94˚C for 10 min. | ||
