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RNA extraction using trizol/tri: Difference between revisions
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→Steps: icons
(→cell lysis: icon) |
(→Steps: icons) |
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=== cell lysis === | === cell lysis === | ||
[[Image:Time required.png]] Cell lysis only takes a few minutes per well, but tissue homogenisation can take 10-20 minutes per sample depending on how tough the tissue is. | |||
* (PBS wash) | * (PBS wash) | ||
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=== phase separation === | === phase separation === | ||
[[Image:Time required.png]] 15-45 min depending on number of samples and whether an additional chloroform wash is necessary | |||
* add chloroform (1/5 volume of trizol; e.g. 0.2ml to 1ml) | * add chloroform (1/5 volume of trizol; e.g. 0.2ml to 1ml) | ||
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</pre> | </pre> | ||
=== RNA precipitation === | === RNA precipitation and wash === | ||
[[Image:Time required.png]] 20-40 min depending on number of samples | |||
* add isopropanol (70% of aqueous phase or 1/2 trizol volume) | * add isopropanol (70% of aqueous phase or 1/2 trizol volume) | ||
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=== RNA wash === | === RNA wash === | ||
[[Image:Time required.png]] 15-30 min depending on number of samples | |||
* wash pellet 70% EtOH (add & vortex briefly) | * wash pellet 70% EtOH (add & vortex briefly) | ||
