5. You know that your inserted gene of interest (GOI) is flanked by EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamH1 restriction endonuclease. In separate reactions, the same preparation was digested to completion with EcoRI and with a mixture of EcoRI and BamH1, respectively. The diagram below shows the appearance of the original molecules and the digestion products in the three digestion mixtures, after separation by electrophoresis in an agarose gel. Lane M - Linear DNA markers of the indicated length in kb Lane 1 - Sample of original (undigested) plasmid DNA prep Lane 2 - Sample of the products of BamH1 digestion Lane 3 - Sample of the products of EcoRI digestion Lane 4 - Sample of the products of digestion of BamH1 + EcoRI a) What was the original DNA molecule's length? b) Was the original plasmid DNA circular or linear? c) How many cuts does the Eco RI make? 1) How many cuts does the Bam HI make? Which map has the correct location of all the restriction enzyme sites of the original plasmid?.

1. 5. You know that your inserted gene of interest (GOI) is flanked by EcoRI sites, which makes
this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has
a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified
DNA preparation of the plasmid is digested to completion with BamH1 restriction endonuclease.
In separate reactions, the same preparation was digested to completion with EcoRI and with a
mixture of EcoRI and BamH1, respectively. The diagram below shows the appearance of the
original molecules and the digestion products in the three digestion mixtures, after separation by
electrophoresis in an agarose gel. Lane M - Linear DNA markers of the indicated length in kb
Lane 1 - Sample of original (undigested) plasmid DNA prep Lane 2 - Sample of the products of
BamH1 digestion Lane 3 - Sample of the products of EcoRI digestion Lane 4 - Sample of the
products of digestion of BamH1 + EcoRI a) What was the original DNA molecule's length? b)
Was the original plasmid DNA circular or linear? c) How many cuts does the Eco RI make? 1)
How many cuts does the Bam HI make? Which map has the correct location of all the restriction
enzyme sites of the original plasmid?