Difference between revisions of "Part:BBa P1014:Experience"
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The P1014 cassette conveys resistance to gentamicin at 20 ug/ml or more in high copy pSB1xx plasmids. | The P1014 cassette conveys resistance to gentamicin at 20 ug/ml or more in high copy pSB1xx plasmids. | ||
| − | This is an image of a growth test plate with | + | This is an image of a growth test plate with pSB1AC3-P1014 in rows 1 and 2, and an unrelated pSB1C3 plasmid in row 3. |
Cells were grown in 700 ul of LB with 20 ug/ml of chloramphenicol overnight, shaking at 37C. Row 1 has 2x serial dilutions of gentamicin, starting at 400 ug/ml on the left. Row 2 has 2x serial dilutions of kanamycin starting at 400 ug/ml on the left. Row 3 has 2x serial dilutions of Gentamicin starting at 400 ug/ml on the left. The rightmost wells have no gentamicin or kanamycin. Growth starts at 12.5 ug/ml in gentamicin, and at 1.6 ug/ml in kanamycin. Growth is inhibited in a non-resistant strain by less than 0.4 ug/ml of gentamicin. A gentamicin concentration of 4 ug/ml allows strong selection. Slightly higher concentrations might be appropriate if trying to distinguish gentamicin resistance from kanamycin resistance. | Cells were grown in 700 ul of LB with 20 ug/ml of chloramphenicol overnight, shaking at 37C. Row 1 has 2x serial dilutions of gentamicin, starting at 400 ug/ml on the left. Row 2 has 2x serial dilutions of kanamycin starting at 400 ug/ml on the left. Row 3 has 2x serial dilutions of Gentamicin starting at 400 ug/ml on the left. The rightmost wells have no gentamicin or kanamycin. Growth starts at 12.5 ug/ml in gentamicin, and at 1.6 ug/ml in kanamycin. Growth is inhibited in a non-resistant strain by less than 0.4 ug/ml of gentamicin. A gentamicin concentration of 4 ug/ml allows strong selection. Slightly higher concentrations might be appropriate if trying to distinguish gentamicin resistance from kanamycin resistance. | ||
Revision as of 18:44, 2 July 2010
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Applications of BBa_P1014
The P1014 cassette conveys resistance to gentamicin at 20 ug/ml or more in high copy pSB1xx plasmids.
This is an image of a growth test plate with pSB1AC3-P1014 in rows 1 and 2, and an unrelated pSB1C3 plasmid in row 3. Cells were grown in 700 ul of LB with 20 ug/ml of chloramphenicol overnight, shaking at 37C. Row 1 has 2x serial dilutions of gentamicin, starting at 400 ug/ml on the left. Row 2 has 2x serial dilutions of kanamycin starting at 400 ug/ml on the left. Row 3 has 2x serial dilutions of Gentamicin starting at 400 ug/ml on the left. The rightmost wells have no gentamicin or kanamycin. Growth starts at 12.5 ug/ml in gentamicin, and at 1.6 ug/ml in kanamycin. Growth is inhibited in a non-resistant strain by less than 0.4 ug/ml of gentamicin. A gentamicin concentration of 4 ug/ml allows strong selection. Slightly higher concentrations might be appropriate if trying to distinguish gentamicin resistance from kanamycin resistance.
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