This experiment was very successful as it was possible to construct a credible restriction map for the unknown plasmid. As part of this experiment, both single digests and double digests consisting of three restriction endonucleases were used to map the restriction sites of the enzymes that make up an unknown plasmid. To separate the DNA fragments based on their distinct number of base pairs, agarose gel electrophoresis had to be performed. As part of gel electrophoresis, a 1kB ladder should be run in the first well. This scale contains numerous known base pair lengths and is run alongside an unknown product to approximate the sizes of the unknown fragments by simply comparing the unknown fragments to coincident fragments of the known scale. This scale gives us the ability to draw precise and accurate conclusions about results derived from gel electrophoresis as it serves as an essential reference point. Thanks to the known fragments in the scale, we were able to create a standard curve. Within the standard curve, the distance traveled by the fragments was plotted against the length of known base pairs within the scale. Once the points were plotted, a line of best fit was constructed and an equation of the line was electronically derived. By inserting the measured distance relative to the distance the fragments travel, shown by “x,” into the equation for the line of best fit, it was possible to calculate the lengths of the base pairs created by the restriction enzymes. In the final step of mapping restriction sites on an unknown plasmid, it was essential to map the individual digest maps first. Individual digest maps were created using both the number of fragments produced... half the paper... lasmids have the ability to survive, and multiple numbers as they expand and reproduce. Furthermore, restriction enzymes have led to important discoveries in the field of cloning. Essentially, because these restriction enzymes allowed the removal of a DNA fragment and its placement in another location, this idea led to scientists being able to integrate exogenous DNA into natural plasmids which could eventually lead to the cloning of plasmid vectors . These plasmids therefore have the ability to self-replicate (neb.com). Discoveries relating to these restriction enzymes paved the way for DNA cloning. Furthermore, DNA mapping is a practical application arising from restriction analysis that now allows scientists to detect insertions and deletions, single nucleotide polymorphisms, and identify genetic disorders (neb.com)