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TOP10 chemically competent cells: Difference between revisions
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→Preparing seed stocks
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===Preparing seed stocks=== | ===Preparing seed stocks=== | ||
* | * Streak TOP10 cells on an [[SOB]] plate and grow for single colonies at 23°C | ||
** room temperature works well | ** room temperature works well | ||
* Pick single colonies into 2 ml of SOB medium and shake overnight at 23 C | * Pick single colonies into 2 ml of SOB medium and shake overnight at 23°C | ||
** room temperature works well | ** room temperature works well | ||
* Add glycerol to 15% | * Add glycerol to 15% | ||
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* Place tubes into a zip lock bag, immerse bag into a dry ice/ethanol bath for 5 minutes | * Place tubes into a zip lock bag, immerse bag into a dry ice/ethanol bath for 5 minutes | ||
** This step may not be necessary | ** This step may not be necessary | ||
* Place in -80 freezer indefinitely. | * Place in -80°C freezer indefinitely. | ||
===Preparing competent cells=== | ===Preparing competent cells=== | ||
