Lan Report - Dna Biochem

(PCR), which isolates small fragments of DNA that have a high degree of variability from

Each human being has something called DNA. DNA is described as genetics and an extremely long macromolecule that is the main component of chromosomes and is the material that transfers genetic characteristics in all life forms. DNA constructs of two nucleotide strands coiled around each other in a ladder like arrangement with the sidepieces composed of alternating phosphate and deoxyribose units and the rungs composed of the purine and pyrimidine bases adenine, guanine, cytosine, and thymine. Each chromosome consist of one continuous thread-like molecule of DNA coiled tightly around proteins and contains a portion of the 6,400,000,000 basepairs that make up your DNA.

PCR permits the synthesis of millions of copies of a specific nucleotide sequence in a few hours. It can amplify the sequence, even when the targeted sequence makes up less than one part in a million of the total initial sample. Steps of the PCR cycle are shown in below figure.

Tierra Harris Cell Biology Lab Report 2 October 12, 2016 Tuesday 10am Dr. Howard DNA Isolation Lab Introduction What is DNA isolation? DNA isolation is the capturing of the personal DNA and that will be isolated in the standard agarose DNA gels. When doing this experiment the student has to understand the meaning of DNA, which is a nucleic acid that contains the genetic instructions for the development and function of living things. DNA is a long polymer which contains the makeup of Adenine (A), Thymine (T), Cytosine (C), and Guanine (G). Within this experiment the student will be using the UV light after finished with the agarose gel to determine Before conducting this experiment I hypothesize that all the DNA results would come out

After the replication fork has been established the strands of DNA are ready for the next stage. On each strand is a

3) Copy: DNA Polymerase joins individual nucleotides with their complementary counterparts to form a new strand of DNA. Replication runs in the 5’ to 3’ direction.

The DNA being heated to 95° C. This will result in the weak hydrogen bonds that clutch the DNA strands together in a helix to break apart. This causes the separation of the strands resulting in the generation of single stranded DNA.

What does the PCR do? PCR is a method by which fragments of DNA can be duplicated. This makes PCR more sufficient amongst other Felds in forensic science.

The primary purpose is to identify a genetic marker or study the function of a specific gene. There are three steps involved in this process which are as follows: denaturation, annealing and elongation. Denaturation involves heating the DNA to agitate the hydrogen bonds, and annealing allows the temperature to be lowered so that the primers can be “annealed” to the single-stranded DNA template. The last step requires DNA polymerase to synthesize a new strand of DNA that is complementary to the RNA strand in the 5’ to 3’ direction (Amplifying DNA: The Polymerase Chain Reaction, 2016). The forward and reverse primers are needed to start the replication process by providing the appropriate nucleotides to the new strand. On the contrary, sanger sequencing makes copies of a target DNA, and the the DNA strand that will be sequenced is separated into two strands, so they can be copied through chemically altered bases. The altered bases cause the process of copying to terminate each time a particular letter is added to the growing DNA chain, which happens to all four bases until the fragments are put together to reveal the original sequence of the original DNA. The aforementioned processes are thoroughly explained to give an overview of the steps involved in providing the end products of the experiment, so an individual can manually decipher

Analysis of DNA from practicals 1 and 2 using the technique of agarose gel electrophoresis and analysis of transfomed E. coli from practical 2 (part B)

Polymerase chain reaction (PCR) is the most common form of DNA analysis (National Institute of Justice). PCR was invented in 1983 by Dr. Kary Mullis. Mullis was working on a project and could not complete it. He said he needed a way to increase the concentration of a specific gene of interest (Oswald, 2017). As he was driving home, he thought that "Using two opposed primers, one complimentary to the upper strand and the other to the lower, then performing multiple cycles of denaturation, annealing and polymerization he could amplify the piece of DNA between the primers" (Oswald, 2017). This is how PCR came about. According to Garibyan and Avashia (2013), "PCR is a simple, enzymatic assay, which allows for the amplification of a specific DNA fragment from a complex pool of data" (p. 1). PCR is a good technique to use because only trace amounts of DNA are needed to generate enough copies to be analyzed. The presence of template DNA, nucleotides, primers, and DNA polymerase are required for every PCR test. It is important to understand what these components are. There are four nucleotides found in DNA and they are adenine, thymine, cytosine, and guanine. Primers are important because this specifies the exact DNA product that must be amplified. Primers are short DNA fragments that have a defined sequence that is complementary to the target DNA that is to be detected

DNA is wound in a double helix structure because of an attraction of charges. Helicase breaks the hydrogen bonds that hold the base pairs, adenine and thymine, guanine and cytosine, together. The separation of the two single strands of DNA forms a Y shape, known as a replication fork. These two separated strands act as a guide for the new strands to form.

The PCR machine is vital to these processes because of its ability to change the temperature based on the step of the process. Step 1 is the initial denaturation at around 94 degrees Celsius where the DNA strands are split into two complementary strands and the Taq DNA polymerase is activated. The next step is annealing at around 62 degrees Celsius in which hydrogen bonds are created between the primers and the top and bottom strands of DNA on opposing ends. During the final extension step at around 72 degrees Celsius, Taq DNA polymerase synthesizes a new DNA strand behind the primers. This three step process is repeated until the desired number of rounds when the samples are held at 4 degrees Celsius to limit Taq

The last stage of the process, joining, involves bonding of complementary nucleotide to each other so as to form new strands. The nucleotides are joined to one another by hydrogen bonds to form a new DNA molecule. This joining continues until a new polynucleotide chain has been formed alongside the old one, forming a new double-helix molecule. This stage of the process also takes place with the assistance of enzymes. The DNA polymerase links the complementary nucleotides

The first is to denature dsDNA through heating to ~96 °C. This separates the two strands of DNA. The exact temperature to be used can be calculated with Tm = 4oC x (no. of G & C) + 2oC x (no. of A & T). Tm is the melting point of the strands and to supply the number of G, C, A, & T ‘s the primer is used.