Question

Biology 107 Lab Assignment 4: Plasmid ID 2.0 1.5 12 0.9 07 05 3000bp 1000bp 500bp 03 02 0.1 Molecular Welght Marker used for gels Reference gel to use for question 6 Hind II (1234) pGFPuv (3337 bp) pHSG (2673 bp) Hind III (234) o49 Hind IlII (1734) Plasmid maps for pGFPuv and pHSG

Answer 1

b. The size of the uncut culture A band in the gel is near 3000 bp. But the HindII cut culture A shows only one band which is little above 3000bp. On comparing the plasmid maps provided, the plasmid pGFPuv has the size of 3337 bp and has a single restriction site of Hind II which will result in a single band on the gel. Hence the pGFPuv plasmid is isolated from the culture A.

c. The size of the uncut culture B band in the gel is near 3000 bp. But the HindII cut culture B shows 2 bands, one below 3000bp and other near 500 bp. On comparing the plasmid maps provided, the plasmid pHSG has the size of 2673 bp and has 2 restriction sites of Hind II which will result in a 2 bands on the gel. Hence the pHSG plasmid is isolated from the culture B.

d. There is a single band in both the uncut and Hind II digested lane of the culture A. The bands are not curved and they are straight. Hence the plasmid in culture A is relaxed.

e. The band in the uncut culture B looks like a curve. Hence the plasmid is supercoiled. The Hind II digested culture B has 2 bands. The band little below 3000 bp looks like a curve and that is supercoiled and the other one near 500 bp looks straight and hence it is linear or relaxed.

a.