Materials and Methods
Restriction enzyme digestion is a method used to identify any differences in DNA sequences among people, the use of restriction enzymes are what help to determine the order of DNA in which they attach to recognition sites and cut the DNA. People with different genes have different recognition sites for restriction enzymes. (Upadhyaya 45-46) In this experiment, four test tubes were used for restriction enzyme digestion reactions which were all filled using a micropipette with 10μl each. Test tubes one and two belong to suspect 1 and test tubes three and four belong to suspect 2. Next, each test tube was filled accordingly with either DNA 1 or DNA 2 with enzymes 1 and 2, see chart below: (Upadhyaya 49)
When transferring…show more content… Then, the following steps called for casting and loading a gel for electrophoresis. In preparation of casting a 50 mL 0.8% gel, the calculated amount of agarose used was 0.4g/50mL. The agarose was added to a 250mL flask containing 50mL of 1xTBE buffer which was swirled in order to rid of clumps. Next, the flask was covered with plastic wrap and microwaved for about a minute in order to dissolve the agarose. When the agarose solution had cooled enough to hold TA: Hien added ethidium bromide which is responsible for latching itself onto DNA in order to make itself detectable in UV light, this was the part where the gel was stained. (Upadhyaya 53) Then the agarose solution was poured into a gel tray that contained a gel comb that is responsible for making the wells. Once the solution was solidified the comb was removed and the gel tray with the gel on it was positioned in a manner so that the row of wells were at the…show more content… As for suspect 1, although, had its DNA with enzyme 1 match up perfectly with CS1 lead to the suspicion that he/she committed the crime but than his/her DNA was mixed with enzyme 2 lead to the conclusion that suspect 1 had no connection with CS2 which is why this suspect is not at fault. However, suspect 2 could in fact be innocent because the suspects DNA could have appeared at that particular area for many reasons. For example, if there was auto theft at a car dealership and suspects’ 2 DNA just so happened to be there because he/she was shopping, keeping in mind that the suspect was at the dealership before the crime was committed which leads to the fact that suspect 2 is innocent. Another reason that suspect 2 could be tied up to the crime is the mere fact that he/she has the same DNA profile as the criminal, it could be read as a false