The following electrophoresis gel resulted from a study of the effects of 3 rest

Q: The following electrophoresis gel resulted from a study of the effects of 3 rest

The approximate size of plasmid pNEW is 4.4 kb because Restriction enzyme Fragment size(kb) Total size (kb) Pst I, Eco RI 3.6, 0.8 4.4 Pst I, Sal I 3.2, 1.2 4.4 Sal I, Eco RI 4, 0.4 4.4 Restriction map Restriction map of pNEW EcoR I 0.8kb 0.4kb Sal I Pst I 3.6kb Restriction enzyme Fragment size(kb) Total size (kb) Pst I, Eco RI 3.6, 0.8 4.4 Pst I, Sal I 3.2, 1.2 4.4 Sal I, Eco RI 4, 0.4 4.4

ID: 307099 • Letter: T

Question

The following electrophoresis gel resulted from a study of the effects of 3 restriction endonucleases (Pst1, EcoR1 and Sal1) both alone and in combination on the plasmid pNEW. Labels above the gel indicate what went into each well; numbers to the right are the estimated sizes, in kilobases (kb), of the fragments comprising each of the bands. Your objective is to use these data to construct a restriction map for pNEW by following the steps presented.

17. Circle the Sall site that would also produce the fragments expected for EcoR1 and Sal 1 (see the bands in lane 6)?

Pan EcoRI SalEsoRl Sal Sal 4.4 kb 4.0kb 3.6kb 3.2kb 1.2kb 0.8kb 0.4 kb bbb kkk 4062 284 4433 100 F

Explanation / Answer

The approximate size of plasmid pNEW is 4.4 kb

because

Restriction enzyme

Fragment size(kb)

Total size (kb)

Pst I, Eco RI

3.6, 0.8

4.4

Pst I, Sal I

3.2, 1.2

4.4

Sal I, Eco RI

4, 0.4

4.4

Restriction map

Restriction map of pNEW EcoR I

0.8kb 0.4kb Sal I

Pst I

3.6kb