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DNA Fingerprinting process

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Introduction

INTRO DNA Fingerprinting in other words DNA profiling is a technique that uses only samples of individual?s DNA to differentiate them from others. Based on the fact that every individual?s DNA profile is unique, the process of DNA fingerprinting is done to acquire the specific DNA profile of the individual. DNA Fingerprinting could be applied to Forensic Science to solve crime, Genetic Screening to identify genetic disorders or hereditary screening to identify individuals that are genetically related. PCR Once the DNA is collected from the individual and extracted from the cell, with breaking the cell membrane and the nuclear membrane by using detergent, the DNA is photocopied using the Polymerase Chain Reaction Technique. The specific sequence of the DNA molecule that is to be replicated must be identified beforehand. A pair of primers ? DNA sequences that are 18-25 base pairs long and are specifically complementary to the DNA sequence identified- are required to be put together in order to mark the chunk of DNA that is to be copied. Why is PCR done? It is done to allow a very large number of human insulin genes available to work with; because when the recombinant plasmids (at the end of the process) ...read more.

Middle

DNA Fragmentation Restriction enzymes (specifically known as restriction endonucleases) function to cut strands of DNA at certain sites. These sites are called restriction sites and they are palindromic; meaning, that the nucleotides are read the same on the forward strand as it is read on the reverse strand. Genetic Engineering- Recombinant DNA Technology is based on the enjoinment of the sticky ends cut by different restriction enzymes when the base pairs are complementary to each other. How do Restriction Enzymes Work? 1. Initially, it cuts from two areas; one cut is through each strand, resulting in the separation of the sugar phosphate backbones and the other cut breaks the phosphodiester bond. 2. Some restriction enzymes form blunt ends by cutting across both the two strands. However most enzymes form sticky ends by cutting the two strands staggered. Sticky ends are the areas located on the DNA which there is an overhang, meaning bases are exposed among the forward and backward ends of the DNA sequence which had been cut by the restriction enzyme. 3. These sticky overhangs are named cohesive ends and they enable the pieces of DNA to recombine with the other cohesive ends when the base pairs on the cohesive ends are complementary to each other; phosphodiester bonds are reformed by DNA ligase. ...read more.

Conclusion

The gel is put inside a buffer as it conducts the electricity which is needed for the electric field to carry out its process without being interrupted; and it controls the pH which is an important factor towards the stability of the DNA molecules. 2. The buffer can be either the TAE (Tris-acetate-EDTA) or TBE (Tris-borate-EDTA); TAE easily becomes exhausted due to its lower buffer capacity than TBE, however the double stranded DNA is able to run much quicker in TAE. 3. An electrophoresis chamber and power supply is essential for the procedure to take place as it aids in the process of separating the DNA sample; giving a low voltage is the best to use because it allows a slower and more precise separation of the DNA sample. 4. The loading buffers used to inject the DNA fragments into the wells are made of a dense liquid solution? e.g glycerol- in order to keep the sample inside the wells and not float away. 5. It also contains a tracking dye that helps with the monitoring of how far the sample has travelled in the gel. ...read more.

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