Dna Synthesis Lab Report

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Introduction:
DNA extraction, PCR, and gel electrophoresis were used to determine the sex of a diecious plant: Silene latifolia. Unlike most plants that are hermaphroditic, diecious plants are either male (XY) or female (XX). The presence of the Y chromosome in a male diecious plant presented a point in which the DNA was be isolated, amplified, and used to differentiate between plant sexes. To identify the sexes three techniques were divided among a couple of weeks.
The first was DNA extraction. DNA extraction worked by opening the cell, adding buffers/ inhibitors to prevent degrading components, and essentially isolating the DNA (Skvirsky, 2015). The second part of the process included amplifying the DNA through PCR (Polymerase Chain Reaction). …show more content…
Throughout, new pipettete tips were used for each sample to prevent cross contamination. There was no need to dilute the DNA prior to placing them in the new tubes. A cocktail was made to place into each tube. The cocktail consisted of 10.5 ul of primer and primer 2 (already added), 47.6 ul of sterile water, and 35 ul of 5 X PCR green flexi buffer, 17.5 of dNTPs and MgCl2, and Tad polymerase added by the instructor. 20 ul of the cocktail was added to each of the 6 tubes and centrifuged for 10 seconds. The tubes were then placed into a thermocycler for multiple heating/cooling rounds including 94℃ denaturing, 60℃ primer annealing and 72℃ DNA polymerization. The samples were left until the following session. The final session included making the agarose mini gel and filling the electrophoresis with 1X TBE buffer. The wells were made to be no more than 2.5 cm from the bottom to the top. 20 ul of each sample was placed in the corresponding wells: 1-positve control, 2-known male, 3-known female, 4-unknown #9, 5-unknown #10, 6-negative control and 7- (10ul) of the ladder. The power source was set to 250V instead of the manual listed 70V and set to run. After several minutes, the gel was removed from the chamber and observed under a UV
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