Polymerase Chain Reaction Lab Report

Polymerase Chain Reaction (PCR) was performed to purify the DNA extract. A mastermix was needed to be made for the PCR products, the mastermix volumes were calculated and shown in table 1. PCR is a simple and inexpensive tool needed to focus on a segment of DNA and a copy it a billion times over. (2) This was needed to purify the DNA samples of the patients which were needed in a gel electrophoresis procedure. The agrose gel electrophoresis process uses electricity to separate DNA fragments by size as they migrate through a gel matrix. (3) Nucleic acid molecules are separated by applying electric field to move a negatively charged molecule through the agrose gel towards the positive charge. (3) The shorter the molecule the faster it travels compared to the larger molecules, because smaller molecules migrate more easily through the pores of the gel. (3) This accurately

(8) It allows us to sequence DNA and RNA much more quickly and cheaper than other techniques such as Sanger sequence. (8) It enables rapid sequencing of large stretches of DNA base pairs of entire genomes with instruments that are able to produce hundreds of gigabases of data in a single sequencing run, enabling researches to form full data sets in a matter of hours or days. (8) How this is done could be explained using an example of gDNA. As you can see in figure 2, the gDNA samples are first fragmented into small segments as shown on picture B, these can be accurately sequenced in millions of parallel reactions. (8)The newly identified bases also known as reads were then reassembled using a known reference as a scaffold which is shown clearly in picture C of figure 2. (8)The aligned reads are then used to reveal the entire sequence of each chromosome shown in picture D. Since the NGS can multi- plex (massive parallel sequence) it saves time to data for multi- sample