Gel Electrophoresis Lab Report

The two techniques that were used to create a DNA profile in this experiment were PCR and gel electrophoresis. The PCR is used to amplify the several DNA samples and gel electrophoresis is performed to separate the DNA fragments according to their size. [6] In the first part of the experiment, PCR amplification of the DNA templates was performed and the products obtained were used to perform gel electrophoresis. The process of PCR allows for the amplification of the DNA samples and the components needed to perform PCR are template DNA, DNA polymerase, primers, buffer, magnesium and nucleotides (dNTPs or deoxy nucleotide triphosphates). [7] DNA samples were added to a master mix, which provides the key ingredients, which are necessary for performing PCR. These components are premixed and optimized and is used in order to simply the process of PCR. [8] The master mix used in this experiment is the Qiagen Master Mix that includes the PCR buffer, MgCl2, and ultrapure dNTPs at optimized concentrations. In order to complete the process of PCR, apart from the master mix only the primers and template has to be added. This method allows
Hence in order to perform several tests many copies of the sample is required, therefore the DNA needs to be amplified. The process of PCR occurs over 35 cycles; each cycle consists of 3 steps involving denaturation, annealing and primer extension. Denaturation is performed at 94˚C for 2 minutes. This step is performed in order to separate the double stranded DNA template into single strands. Next, annealing is performed usually between 42-65˚C for 15-60 s. In this step, the reason for the reduced temperature is so that the primers can anneal with the denatured DNA and also acts as primers for the DNA polymerase. Lastly, extension occurs at 72˚C for 10 minutes where the DNA strands synthesize via DNA polymerase.