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Science Articles How does DNA Fingerprinting Work? and what can it tell us about wine. People everywhere expected the new millennium to bring surprises. But the particular shock and horror that rippled through the international viticulture community in 2000 was most unexpected. It had been found that sixteen of the most highly prized varieties of wine-making grapes were the products of mating between the classic Pinot and the classically undervalued Gouais grape. This blew the proverbial cork off the industry's bottle because the Gouais was considered such an inferior specimen that there were even attempts to ban its cultivation in France during the Middle Ages. This proves that humble origins can still produce superior quality. More practically, though, knowledge of their heritage allows improved breeding of highly desirable subspecies of grape. And viticulturists everywhere had DNA fingerprinting technology to thank. DNA fingerprinting is a term that has been bandied about in the popular media for many years, largely due to its power to condemn and save, but what does it involve? In short, it is a technique for determining the likelihood that genetic material came from a particular individual or group. 99% of human DNA is identical between individuals, but the 1% that differs enables scientists to distinguish identity. In the case of the grapes, scientists compared the similarities between different species and were able to piece together parent subspecies that could have contributed to the present prize-winning varieties. The DNA alphabet is made up of four building blocks – A, C, T and G, called base pairs, which are linked together in long chains to spell out the genetic words, or genes, which tell our cells what to do. The order in which these 4 DNA letters are used determines the meaning (function) of the words, or genes, that they spell. But not all of our DNA contains useful information; in fact a large amount is said to be “non-coding” or “junk” DNA which is not translated into useful proteins. Changes often crop up within these regions of junk DNA because they make no contribution to the health or survival of the organism. But compare the situation if a change occurs within an essential gene, preventing it from working properly; the organism will be strongly disadvantaged and probably not survive, effectively removing that altered gene from the population. For this reason, random variations crop up in the non-coding (junk) DNA sequences as often as once in every 200 DNA letters. DNA fingerprinting takes advantage of these changes and creates a visible pattern of the differences to assess similarity. Stretches of DNA can be separated from each other by cutting them up at these points of differences or by amplifying the highly variable pieces. „Bands‟ of DNA are generated; the number of bands and their sizes give a unique profile of the DNA from whence it derived. The more genetic similarity between a person – or grape – the more similar the banding patterns will be, and the higher the probability that they are identical. In the non-coding regions of the genome, sequences of DNA are frequently repeated giving rise to so-called VNTRs - variable Left - DNA fingerprints from 6 different people, 1 in each lane (column). DNA can be cut into shorter pieces by enzymes called "restriction endonucleases". The pieces of DNA can then be separated according to their size on a gel. Each piece of DNA forms a band (the white lines on the gel). The smallest pieces travel the furthest and are therefore closest to the bottom of the gel. The larger pieces travel shorter distances and are closer to the top. number tandem repeats. The number of repeats varies between different people and can be used to produce their genetic fingerprint. In the simple example shown above, person A has only 4 repeats whilst person B has 7. When their DNA is cut with the restriction enzyme Eco RI, which cuts the DNA at either end of the repeated sequence (in this example), the DNA fragment produced by B is nearly twice as big as the piece from A, as shown when the DNA is run on a gel (right). The lane marked M contains marker pieces of DNA that help us to determine the sizes. If lots of pieces of DNA are analysed in this way, a 'fingerprint' comprising DNA fragments of different sizes, unique to every individual, emerges. But why bother? After all, I know where my wine comes from – Tesco's, right? Well, there are many relevant applications of DNA fingerprinting technology in the modern world, and these fall into three main categories: To find out where we came from, discover what we are doing at the present, and to predict where we are going. In terms of where we came from, DNA fingerprinting is commonly used to probe our heredity. Since people inherit the arrangement of their base pairs from their parents, comparing the banding patterns of a child and the alleged parent generates a probability of relatedness; if the two patterns are similar enough (taking into account that only half the DNA is inherited from each parent), then they are probably family. However, DNA fingerprinting cannot discriminate between identical twins since their banding patterns are the same. In paternity suits involving identical twins - and yes, there have been such cases - if neither brother has an alibi to prove that he could not have impregnated the mother, the courts have been known to force them to split child care costs. Thankfully there are other, less “Jerry Springer-esque”, applications that teach us about our origins. When used alongside more traditional sociological methodologies, DNA fingerprinting can be used to analyse patterns of migration and claims of ethnicity. DNA Fingerprinting can also tell us about present-day situations. Perhaps best known is the use of DNA fingerprinting in forensic medicine. DNA samples gathered at a crime scene can be compared with the DNA of a suspect to show whether or not he or she was present. Databases of DNA fingerprints are only available from known offenders, so it isn't yet possible to fingerprint the DNA from a crime scene and then pull out names of probable matches from the general public. But, in the future, this may happen if DNA fingerprints replace more traditional and forgeable forms of identification. In a real case, trading standards agents found that 25% of caviar is bulked up with roe from different categories, the high class equivalent of cheating the consumer by not filling the metaphorical pint glass all the way up to the top. DNA fingerprinting confirmed that the „suspect‟ (inferior) caviar was present at the crime scene. In the example shown on the left, DNA collected at the scene of a crime is compared with DNA samples collected from 4 possible suspects. The DNA has been cut up into smaller pieces which are separated on a gel. The fragments from suspect 3 match those left at the scene of the crime, betraying the guilty party. Finally, genetic fingerprinting can help us to predict our future health. DNA fingerprinting is often used to track down the genetic basis of inherited diseases. If a particular pattern turns up time and time again in different patients, scientists can narrow down which gene(s), or at least which stretch(es) of DNA, might be involved. Since knowing the genes involved in disease susceptibility gives clues about the underlying physiology of the disorder, genetic fingerprinting aids in developing therapies. Pre-natally, it can also be used to screen parents and foetuses for the presence of inherited abnormalities, such as Huntington‟s disease or muscular dystrophy, so appropriate advice can be given and precautions taken as needed. Acknowledgement: This article was co-authored with Dr Chris Smith, who also compiled the images. - May 2004 References Make your own DNA fingerprint DNA From The Beginning Q's and A's about DNA Fingerprinting Link: http://www.thenakedscientists.com/HTML/articles/article/dalyacolumn8.htm/ DNA Fingerprinting in Human Health and Society Written by David F. Betsch, Ph.D., Biotechnology Training Programs, Inc. Edited by Glenda D. Webber, Iowa State University Office of Biotechnology. Issued in furtherance of Cooperative Extension work, Acts of Congress of May 8 and June 30, 1914, in cooperation with the U.S. Department of Agriculture and Cooperative Extension Services of Illinois, Indiana, Iowa, Kansas, Michigan, Minnesota, Missouri, Nebraska, North Dakota, Ohio, South Dakota, and Wisconsin. June, 1994 Like the fingerprints that came into use by detectives and police labs during the 1930s, each person has a unique DNA fingerprint. Unlike a conventional fingerprint that occurs only on the fingertips and can be altered by surgery, a DNA fingerprint is the same for every cell, tissue, and organ of a person. It cannot be altered by any known treatment. Consequently, DNA fingerprinting is rapidly becoming the primary method for identifying and distinguishing among individual human beings. An additional application of DNA fingerprint technology is the diagnosis of inherited disorders in adults, children, and unborn babies. The technology is so powerful that, for example, even the blood-stained clothing of Abraham Lincoln could be analyzed for evidence of a genetic disorder called Marfan's Syndrome. The Structure of DNA The characteristics of all living organisms, including humans, are essentially determined by information contained within DNA that they inherit from their parents. The molecular structure of DNA can be imagined as a zipper with each tooth represented by one of four letters (A, C, G, or T), and with opposite teeth forming one of two pairs, either A-T or G-C. The letters A, C, G, and T stand for adenine, cytosine, guanine, and thymine, the basic building blocks of DNA. The information contained in DNA is determined primarily by the sequence of letters along the zipper. For example, the sequence ACGCT represents different information than the sequence AGTCC in the same way that the word "POST" has a different meaning from "STOP" or "POTS," even though they use the same letters. The traits of a human being are the result of information contained in the DNA code. Living organisms that look different or have different characteristics also have different DNA sequences. The more varied the organisms, the more varied the DNA sequences. DNA fingerprinting is a very quick way to compare the DNA sequences of any two living organisms. Making DNA Fingerprints DNA fingerprinting is a laboratory procedure that requires six steps: 1: Isolation of DNA. DNA must be recovered from the cells or tissues of the body. Only a small amount of tissue - like blood, hair, or skin - is needed. For example, the amount of DNA found at the root of one hair is usually sufficient. 2: Cutting, sizing, and sorting. Special enzymes called restriction enzymes are used to cut the DNA at specific places. For example, an enzyme called EcoR1, found in bacteria, will cut DNA only when the sequence GAATTC occurs. The DNA pieces are sorted according to size by a sieving technique called electrophoresis. The DNA pieces are passed through a gel made from seaweed agarose (a jelly- like product made from seaweed). This technique is the biotechnology equivalent of screening sand through progressively finer mesh screens to determine particle sizes. 3: Transfer of DNA to nylon. The distribution of DNA pieces is transferred to a nylon sheet by placing the sheet on the gel and soaking them overnight. 4-5: Probing. Adding radioactive or colored probes to the nylon sheet produces a pattern called the DNA fingerprint. Each probe typically sticks in only one or two specific places on the nylon sheet. 6: DNA fingerprint. The final DNA fingerprint is built by using several probes (5-10 or more) simultaneously. It resembles the bar codes used by grocery store scanners. Uses of DNA Fingerprints DNA fingerprints are useful in several applications of human health care research, as well as in the justice system. Diagnosis of Inherited Disorders DNA fingerprinting is used to diagnose inherited disorders in both prenatal and newborn babies in hospitals around the world. These disorders may include cystic fibrosis, hemophilia, Huntington's disease, familial Alzheimer's, sickle cell anemia, thalassemia, and many others. Early detection of such disorders enables the medical staff to prepare themselves and the parents for proper treatment of the child. In some programs, genetic counselors use DNA fingerprint information to help prospective parents understand the risk of having an affected child. In other programs, prospective parents use DNA fingerprint information in their decisions concerning affected pregnancies. Developing Cures for Inherited Disorders Research programs to locate inherited disorders on the chromosomes depend on the information contained in DNA fingerprints. By studying the DNA fingerprints of relatives who have a history of some particular disorder, or by comparing large groups of people with and without the disorder, it is possible to identify DNA patterns associated with the disease in question. This work is a necessary first step in designing an eventual genetic cure for these disorders. Biological Evidence FBI and police labs around the U.S. have begun to use DNA fingerprints to link suspects to biological evidence - blood or semen stains, hair, or items of clothing - found at the scene of a crime. Since 1987, hundreds of cases have been decided with the assistance of DNA fingerprint evidence. Another important use of DNA fingerprints in the court system is to establish paternity in custody and child support litigation. In these applications, DNA fingerprints bring an unprecedented, nearly perfect accuracy to the determination. Personal Identification Because every organ or tissue of an individual contains the same DNA fingerprint, the U.S. armed services have just begun a program to collect DNA fingerprints from all personnel for use later, in case they are needed to identify casualties or persons missing in action. The DNA method will be far superior to the dogtags, dental records, and blood typing strategies currently in use. For Further Reading "DNA fingerprints witness for the prosecution." Discover. June 1988, p. 44. DNA Identity Testing Information Package. Available from LifeCodes, Inc., Stamford, Connecticut. Phone toll-free: 1 (800) 543-3263. Genetic Witness Forensic Uses of DNA Tests. U.S. Office of Technology Assessment. July 1990. Phone: (202) 224-8996. "Molecular advances in genetic disease." Science. May 8, 1992. "The promise and pitfalls of molecular genetics." Science. July 10, 1992. Link: http://www.accessexcellence.org/RC/AB/BA/DNA_Fingerprinting_Basics.php . Science Articles How does DNA Fingerprinting Work? and what can it tell us about. advances in genetic disease." Science. May 8, 1992. "The promise and pitfalls of molecular genetics." Science. July 10, 1992. Link: http://www.accessexcellence.org/RC/AB/BA/DNA_Fingerprinting_Basics.php