A kanamycin-resistant, ampicillin-resistance plasmid (3 kb) is cleaved with the PstI enzyme, which cuts within the ampicllin resistance gene. The digested plasmid is ligated to a 5-kb PstI DNA fragment from Drosophila (fruit fly). The DNA mixture was then transformed into E.coli host cells.
(i). Which antibiotic would you put into the medium (nutrient agar plate) to select for cells that contain the plasmid?
(ii). Explain step by step, how you would select for cells containing the recombinant Drosophila DNA.
(iii) If different colonies from the plate in part (ii) are cultured in nutrient broth and DNA was isolated from each of the bacterial cultures, what restriction patterns would you expect to see following PstI digestion and agarose gel electrophoresis?
A kanamycin-resistant, ampicillin-resistance plasmid (3 kb) is cleaved with the PstI enzyme, which cuts within the ampicllin resistance gene. The digested plasmid is ligated to a 5-kb PstI DNA fragment from Drosophila (fruit fly). The DNA mixture was then transformed into E.coli host cells.
(i). Which antibiotic would you put into the medium (nutrient agar plate) to select for cells that contain the plasmid?
(ii). Explain step by step, how you would select for cells containing the recombinant Drosophila DNA.
(iii) If different colonies from the plate in part (ii) are cultured in nutrient broth and DNA was isolated from each of the bacterial cultures, what restriction patterns would you expect to see following PstI digestion and agarose gel electrophoresis?
