Difference between revisions of "Part:BBa K1323017:Experience"
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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 ug/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 uL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 ug/mL of erythromycin and chloramphenicol. The negative growth control was 75 ug/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. | 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 ug/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 uL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 ug/mL of erythromycin and chloramphenicol. The negative growth control was 75 ug/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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===User Reviews=== | ===User Reviews=== | ||
Revision as of 04:32, 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 ug/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 uL of the overnight cultures were used to inoculate fresh TSB that was supplemented with 10, 50, or 100 ug/mL of erythromycin and chloramphenicol. The negative growth control was 75 ug/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.
User Reviews
UNIQf9d0250d47b0f909-partinfo-00000000-QINU UNIQf9d0250d47b0f909-partinfo-00000001-QINU