The following electrophoresis gel resulted from a study of t

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

Solution

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