You've Cloned a 2-Kb Fragment of DNA into a Bacterial

Multiple Choice

You've cloned a 2-kb fragment of DNA into a bacterial cloning vector and want to construct a restriction map of the insert. You amplify the 2-kb insert using PCR, purify it, and subject it to differential digestion with the enzymes EcoRI and HindIII, gel-fractionate the digests, and visualize the restriction patterns by staining the gels with ethidium bromide to generate the following results.
-Using these data, indicate the order of restriction sites in the DNA insert. $$\begin{array} { l l } & \text { Restriction fragment lengths } \\\hline\text { EcoRI } & 350 \mathrm { bp } , 500 \mathrm { bp } , 1250 \mathrm { bp } , 100 \mathrm { bp } \\\text { HindIII } & 350 \mathrm { bp } , 650 \mathrm { bp } , 1000 \mathrm { bp } \\\text { EcoRI/HindIII } & 100 \mathrm { bp } , 150 \mathrm { bp } , 200 \mathrm { bp } , 300 \mathrm { bp } , 350 \mathrm { bp } , 900 \mathrm { bp }\end{array}$$

A) EcoRI, HindIII, EcoRI, HindIII, EcoRI
B) HindIII, EcoRI, HindIII, EcoRI, HindIII
C) HindIII, EcoRI, HindIII, HindIII, EcoRI
D) HindIII, EcoRI, EcoRI, HindIII, EcoRI
E) EcoRI, EcoRI, HindIII, EcoRI, HindIII