4. (TCO 9) Provide a detailed description of the techniques used to make a DNA fingerprint. What are some of the uses and applications of DNA fingerprinting? Solution DNA fingerprinting is a technique used to determine the nucleotides sequences of DNA which are unique to each individual. Technique 1. Extraction of the DNA from the source the DNA is extracted from blood sample, hair follicles etc.available sample. 2.DNA is cut into fragments the DNA molecules broken with the help of restriction endonuclease. Here the cleaning is double strand cut producing DNA fragments of different lengths this fragment are also called restricted fragment length polymorphism Manyi of this fragment contain vntr 3. Separation of the fragments using gel electrophoresis. As the DNA molecule is negatively charged hence it will move towards positive or not in the setup the gel based matrix provides tiny pores through which DNA molecules travel the larger molecules travel slowly where is the smallest mens travel quickly from the loading point at the end of the experiment DNA pieces of equal length obtained. 4. The DNA fragments or now treated with alkaline chemicals to facilitate denaturation into single stranded DNA this is very important step. 5. Southern blotting technique in this technique nitrocellulose membrane is used the DNA is bloated on suitable membrane like nitrocellulose or nylon membranes as they have good binding capacity the membrane is subjected to gentle pressure due to this single stranded DNA fragments are pulled and transfer onto the membrane . the membrane contains replica of the DNA. Hybridisation with suitable DNA probe which is single stranded DNA having complementary sequence to the desired DNA. Before using the probe the DNA of tagged with fluorescent dyes to help in detection of the desired DNA excess probea are washed away. 5. the DNA sample is visualised using autoradiography the hybridisation pattern is called DNA fingerprint having a sequence complementary to the probe. 6. PCR technique is a technique is useful to synthesise millions of copies of the DNA sequence when low amount of DNA is available for the study this technique is used modifications of PCR technique like r a p d PCR rflp PCR helps in giving accurate results. Applications of DNA fingerprinting : 1.This test is used in the case of disputes regarding paternity testing . 2 it is useful tool in forensic applications 3.It is used to assess migration pattern of ancient population 4.it is used to determine Genetic diversity is in the evolutionary biology. 4. It is used to diagnose inherited disorders in both prenatal and newborn babies examples huntington\'s disease Alzheimer\'s Sickle Cell anaemia Thalassemia haemophilia. 5. DNA fingerprinting is used to come from confirm cell line identity in a cell line collection. 6. It also helps in developing cures for inherited disorders..