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Transforming bacteria: Difference between revisions
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===Competent cells:=== | ===Competent cells:=== | ||
1) Innoculated a 5ml LB culture in 15ml tube with a DH5α colony. Grown overnight in 37°C incubator. Not shaking (might be better if it was). | 1) Innoculated a 5ml LB culture in 15ml tube with a DH5α colony. | ||
Grown overnight in 37°C incubator. Not shaking (might be better if it was). | |||
2) Transferred the 5ml O/N culture into an 'autoclaved' conical flask containing 100ml LB broth and back into the 37°C incubator for about an hour. Then onto hot plate/,magnetic stirrer with a water bath on top to buffer the temperature a bit, it seemed easiest to keep the temperature stable at around 34 to 36°C, not ideal but it worked this time. | 2) Transferred the 5ml O/N culture into an 'autoclaved' conical flask containing 100ml LB broth and back into the 37°C incubator for about an hour. Then onto hot plate/,magnetic stirrer with a water bath on top to buffer the temperature a bit, it seemed easiest to keep the temperature stable at around 34 to 36°C, not ideal but it worked this time. | ||
3) Cells allowed to grow until cloudy (much cloudier than last time), it would be nice to be able to quantify this! The closest I can get is to say that you can only see the magnetic stirring flea when it's on the near side of the flask, so almost opaque. This took about 3 hours. | 3) Cells allowed to grow until cloudy (much cloudier than last time), it would be nice to be able to quantify this! The closest I can get is to say that you can only see the magnetic stirring flea when it's on the near side of the flask, so almost opaque. This took about 3 hours. | ||
