Difference between revisions of "Part:BBa K1323017:Experience"
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==Experience== | ==Experience== | ||
| − | The construct (pSB1CE3 shuttle vector) was electroporated into | + | The construct (pSB1CE3 shuttle vector) was electroporated into ''S.epidermidis''. Three colonies were streak purified and diagnostic restriction digests were done to confirm that the insert was present. Overnight TSB cultures were supplemented with 10 µg/mL erythromycin, and the inoculation was done in triplicate. The negative control for this experiment was ''S.epidermidis'' (ATCC12228), it lacks erythromycin resistance but has chloramphenicol resistance. 10 µL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 µg/mL of erythromycin and chloramphenicol. The negative growth control was 75 µg/mL of kanamycin and the positive growth control was TSB without antibiotic. The samples were incubated at 37 ͒C, shaking at 200 rpm. The OD600 was read after 6 hours of incubation. |
Revision as of 05:17, 17 October 2014
Experience
The construct (pSB1CE3 shuttle vector) was electroporated into S.epidermidis. Three colonies were streak purified and diagnostic restriction digests were done to confirm that the insert was present. Overnight TSB cultures were supplemented with 10 µg/mL erythromycin, and the inoculation was done in triplicate. The negative control for this experiment was S.epidermidis (ATCC12228), it lacks erythromycin resistance but has chloramphenicol resistance. 10 µL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 µg/mL of erythromycin and chloramphenicol. The negative growth control was 75 µg/mL of kanamycin and the positive growth control was TSB without antibiotic. The samples were incubated at 37 ͒C, shaking at 200 rpm. The OD600 was read after 6 hours of incubation.
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