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Lactococcus transformation: Difference between revisions
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1. Grow cells overnight in 25ml of GM17 (350µl glucose solution to 25ml M17).<br> | 1. Grow cells overnight in 25ml of GM17 (350µl glucose solution to 25ml M17).<br> | ||
2. Add 1ml of overnight culture into 25ml SGM17 + 2.0% glycine. <br> | 2. Add 1ml of overnight culture into 25ml SGM17 + 2.0% glycine. <br> | ||
::*(25ml M17 + 350µl glucose solution +5g Sucrose + 0.5g glycine). <br> | |||
3. Grow for ~4 hours until OD600 ~ 0.7. <br> | 3. Grow for ~4 hours until OD600 ~ 0.7. <br> | ||
4. Chill culture on ice for 10 mins. <br> | 4. Chill culture on ice for 10 mins. <br> | ||
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11. Subculture 100µl into 900ul prewarmed M17+ and incubate for 2 hours. <br> | 11. Subculture 100µl into 900ul prewarmed M17+ and incubate for 2 hours. <br> | ||
12. Add entire subculture to GM17 with antibiotic. <br> | 12. Add entire subculture to GM17 with antibiotic. <br> | ||
::*1ug/ml Erm for plates. <br> | |||
::*5ug/ml Erm for culture. | ::*5ug/ml Erm for culture. | ||
