Part:BBa_P1014:Experience

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Applications of BBa_P1014

The P1014 cassette conveys resistance to gentamicin at 12 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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