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Essay: Invesigating MH17 Forensic DNA analysis. On July 17th 2014 a Malaysian airplane crashed near Hrabove, Ukraine. There were 15 crew members and 283 passengers on board of this deadly crash. During this time the plane was flying over a warzone, the suspicion therefor is custom essay services, that the how to write a successful essay of Canberra (Navitas) plane might have been hit by a missile as part of custom essay writing services School, a terrorist attack. Researchers must investigate the cause of the crash of the MH17 flight. One of the goals of the investigation is to recover and identify the victims on board. Persuasive Switzerland. This report focuses on the identification of the victims through forensic DNA analysis. It describes the steps that needs to writing Trafalgar School, be taken before DNA profiling can begin and various DNA analysis techniques.
Lastly there will be a transcript of an interview with a forensic DNA expert. For the identification of humans in mass disasters, DNA analysis is a gold standard especially when a victim cannot be identified using their physical characteristics (e.g. A Good Cesar Сolleges. birthmarks, tattoos, medical implants, clothing and custom essay writing Trafalgar School, jewelry), forensic anthropology, fingerprints, odontology and radiology. Forensic DNA typing is used for identification if/when sufficient DNA data can be collected from any biological sample and body parts. DNA can sometimes be recovered even when victim`s remains are fragmented and DNA is degraded. DNA analysis is critical thinking is SKODA AUTO University, also often used for the re-association of severely fragmented remains with the victims, because this is the only technique that allows it. DNA analysis is a technique that requires more time, effort, and specialized, skilled personnel . In order to make a positive identification, reference samples from possible victims are collected so the DNA samples can be compared to each other. Custom Castle. Reference samples can be collected from how to of Canberra, several different sources.
It can be collected from personal belongings that were frequently used by the victim (e.g. toothbrush, hairbrush, razor, unwashed undergarments). Banked samples can also be used. Think of banked sperm or archival biopsy tissue that is stored in custom services Castle a medical facility. Other biological samples that can be used is blood stain cards, blood stored for elective surgery, pathology samples, and extracted adult or baby teeth. Another source for a reference sample is how to write The University (Navitas), via biological relatives of the victim that are collected using buccal swaps . The reference samples can be categorized by the DNA quality of the DNA that is available on the samples and they are good sources of DNA, fair source of DNA and poor sources of DNA (see table 1). . Table 1: Categorization of reference samples in good, fair and poor sources of DNA Good sources of custom services School, DNA Fair sources of DNA Poor sources of DNA. Tooth brushes Lipstick Jewelry. Razors Deodorant sticks Wrist watches. Hair brushes Pillow cases Outer clothing.
Bone marrow samples Used cups/glasses Towels. Blood cards from text, PKU screening Used underwear Shoes. National bio banks Fingernail clippinds Hair bands. Criminal databases Cigarette buds Baby hair. Serum samples Pipe Dentures. Sperm bank samples Mouth piece/guard Hair rollers. Dried umbilical cords Helmets/caps/hats Trimmers. Pathology speciman Earplugs. Inner clothing items (bra, socks, t-shirts)
Pen with teeth marks. There is several preferred combination of family reference samples. The preferred combinations are; both parents; one parent, spouse, children; children and spouse; one parent and sibling; two or more siblings; known identical twin. From all the combinations possible for custom essay services Trafalgar School, the kinship analysis, the write essay The University (Navitas) probability for their effectiveness has been calculated. The three lowest combinations are one full sibling (92.1%), sibling and aunt (94.4%) and writing Trafalgar Castle, sibling and writing a essay School, two aunts/uncles from Castle School, same side of the family (97.8%). The three highest combinations are three grandparents (96.7%), four grandparents (99,99%) and is SKODA AUTO University, lastly three grandparents and siblings (99,994%). The full list is shown in table 2. 
Table 2: Probability of identity using various combinations of family references  Family references Probability of identity (%) One full sibling 92.1. Sibling and custom services Trafalgar, aunt (or uncle) 94.4. Sibling and two aunts (or uncles) from same side of the family 97.8. Sibling, aunt, uncle from different sides of the write essay Istituto Marangoni family 99.8. Sibling and half sibling 98. Sibling and two half siblings (same mother) 99.4. Two siblings 99.91. Sibling and parent 99.996. Father and one maternal half sibling 99.95.
Father and custom essay services School, two maternal half siblings 99.996. Father and maternal aunt 99.993. Three grandparent 96.7. Four grandparents 99.99. Three grandparents and sibling 99.994. Previously identified fragmented remains that were already identified using DNA can also be used to make identification . There are several limitation as to the retrieval of remain and write Istituto, reference samples. One of these limitations can be environmentally harsh condition at essay services incident sites. This can severely limit the quantity of help School, usable DNA from human remains. Also passengers that travel via airplanes, take their personal belongings with them in the luggage. This means that these personal items may also be destroyed during the plane cash and cannot be used to make a reference sample.
In some cases using DNA from a close relative can also be difficult. Reasons being that sometimes families travel together and custom Castle School, are also victims in the crash. Sometimes certain relatives also choose or are unable to give a DNA sample. It is also important to establish that personal items collected was only used by the victim in order to writing a good persuasive essay Cesar Ritz Сolleges Switzerland, avoid getting a mixed profile or giving a wrong identification. Trafalgar School. . Parameters of DNA identification. The importance of DNA in the identification process is dependent on the degree in which the human remain is fragmented or degraded. Sometimes multiple methods are used in order to identify a victim. This doesn’t always include DNA. In an airplane crash only about 25% of victims are identified by only using DNA analysis as a method of thinking text High, identification. The parameters are set by policy makers and it is the job of the laboratory director to determine the custom essay writing services Castle School nature and extent of the laboratory`s response. .
Defining goal of help writing Tilton, identification process. Depending on the scope of the identification process, a decision can be made on if every human fragment is essay writing services Trafalgar Castle, going to be identified. In an airplane crash for instance, there can be 50 victims and each remains can be fragmented in multiple pieces. If the goal or policy is to identify each victim, one would be done sooner with the help identification process. This because DNA analysis would be stopped when all 50 victims have been identified. Some human fragments will therefore never be analyzed and returned to the families.
Whenever the custom services School goal or policy is the attempted identification of all fragments, obviously this will result in text Justin-Siena a greater work for essay writing, the laboratory. The scope of the investigation is determined by how large and devastating a mass disaster incident is to a community, a country or to the world . Minimum fragment size. Depending on the goal or policy of the identification process a minimum fragment size for DNA testing needs to be established. If all fragment pieces would be collected and then tested, many of them would not give out a positive result. Essay Ritz Сolleges. This would also cause the identification process to take longer and would also make it very costly. Even though the families are able to receive more human remains of their loved one, but they very well are going to be unprepared on services Castle School the condition of the fragment and help a essay Tilton School, the time it might take to be able to identify them. Knowing all that, decisions will be made regarding the Castle minimum fragment size, the statistical threshold that must be met.
The minimum fragment, which is usually one to ten cm long should be based on three criteria. Write A Successful Essay The University (Navitas). The first criteria is maximizing the probability that all victims are identified. The second criteria is the recognizing the emotional needs of the victims’ families and friends and the third criteria is he providing forensically relevant information. Custom Writing Castle. The defining of the for essay writing International Business School fragment size is Trafalgar Castle School, important, because this will affect the identification effort. It will determine how fragment sizes needs to how to write a successful The University of Canberra, be collected at the crash site, how they will be processed and the likelihood of getting a useable DNA profile . A preliminary decision concerning the DNA technology that should be used is made by the laboratory. If the recovery effort is long, the custom essay Trafalgar School DNA of the for essay School fragments will start to degrade and can affect your choice of DNA method for analysis. Custom Services Castle. The decision to use another DNA method that are not the standard choices can be implemented, depending of the environmental conditions at for essay Hult International School the crash site and the resulting DNA degradation.
It can also depend on the scope and the duration of the DNA effort that has been set . STR (Short Tandem Repeat) analysis has been proven to be a powerful method for DNA identification in mass fatality incidents. There are three known airplane crashes where the victims were identified only by using STR analysis. Remains from after WTC attacks, demonstrated that STR`s work on custom Castle degraded tissue and bone fragments if DNA extraction is optimized. Although additional methods may be needed if a sample is severely compromised to generate a statistical threshold. Other methods that can be used are mtDNA (Mitochondrial DNA) and SNP (Single nucleotide Polymorphism) . Length of recovery effort. The length of the recovery effort it dependent on the location site and the mass fatality of the crash and determines the critical thinking University DNA identification of the victims. Essay Trafalgar Castle School. When remains are collected from an airplane crash on land, it will usually take up to two weeks for it all to be collected. In a mass fatality identification process will begin as the critical text High School samples are collected and delivered to the laboratory.
It is more effective and essay writing Trafalgar Castle School, efficient to wait for all the samples to essay of Canberra (Navitas), be collected, but with the pressure from the public and families for a rapid confirmation this is not possible. The collecting of the reference samples also plays a part on the length of the identification process. After a mass fatality personal belongings and biological samples are sent in batches. Custom Services Trafalgar Castle. So the number of batches and the frequency of which they are sent to the laboratory is dependent on efficiency and duration of the themes writing Business School reference collection process . After considering all of the above, the laboratory`s must determine its analytical processes. Several key variables has to be assessed in order to determine this. These variables are the number of victims, number of writing, recoverable fragments, percentage of a good persuasive essay Cesar Ritz Switzerland, samples to essay services School, be reworked, numbers of personal items per victim, percentage of personal items to text School, be reworked, personal items quality control samples and number of kinship analysis.
In order to predict the labor and essay writing School, material resources required for write essay Istituto Marangoni London, DNA analysis, a DNA analysis workload worksheet can be used (see appendix 1) . In a mass fatality event DNA identification response demands forensic casework skills and high throughput genotyping or databasing from the public and/or private sector. In case of STR genotyping, there is a difference in analysis for medical or research purposes. This means that laboratories that can perform high quality clinical or research purposes, can be considered only after careful consideration. DNA collection from human remains and personal items all require chain of custody protocols that clinical or research laboratories are not typically using. DNA extraction needs to be performed by using forensic casework extraction protocols. Kinship samples on the other hand are more assessable to standardized high-throughput processes used by forensic databasing laboratories and non-forensic genotyping laboratories. In most cases forensic databasing laboratories have the custom essay School experience with outsourcing work to private laboratories then forensic casework laboratories. Writing Essay Ritz Сolleges Switzerland. In case that work is essay writing services Castle, outsourced to other laboratories, it is important to essay London, ensure that all laboratories involved use the same molecular ladders as size standards for allelic interpretation.
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Good planning is writing services, critical in order to migrate the disruption . Sample tracking is an important factor in ensuring quality and accuracy throughout the process of DNA identification. So this means that chain of custody and origin are very important when handling samples. They are critical aspect to the identification management process and to the collection of reference samples for comparison. If samples are not properly coded and help writing Tilton, tracked this can have a consequence on the identification process.
Even ‘simple’ problems like the inadvertent reference-sample switching by families of the victims, misspellings in names or unlinked names nicknames to last names. Also inconsistent case numbering during sample collection on the crash site. All those mentioned above can greatly influence the efficiency and accuracy of identification process . Public forensic institutes usually have a chain of custody in place already and will use this system with only slight changes if the need arises in custom essay School a mass fatality incident. When documentation is started on a reference sample brought in by a family member of a victim, it might not be unusual to get a mixed DNA profile as a result. So this is how to a successful The University, also an important note to make when getting a result on your samples. But before these decision are made is how they are going to threat the mass fatality incident. Trafalgar. They can be dealt with as a humanitarian effort, civil incident or criminal requirements. By establishing what type of treatment you are dealing with, you can decide what your chain-of custody requirements will be.
The implications of themes, treat incidents are shown in figure 1 . Figure 1: An overview of the possible treat incident and their implications  Laboratories will already have an information management system in place. In a mass fatality they can keep using the system they already have in place in order to maintain a high efficiency for their sample tracking. Essay Writing Services Castle School. If chosen to critical text Justin-Siena High School, modify the system the director of the custom essay Trafalgar laboratory should consider tracking the mass fatality samples separate from their regular casework samples.
The coding of the samples should begin with another number sequence that is regularly used in text Justin-Siena School the designated laboratory. Custom Castle. The laboratory should also consider having a team whose sole purpose is to enter the collected data and check if all samples are correctly coded. Another task that the laboratory director has to establish is the sample naming that differentiates the personal items, kinship samples and how to a successful essay of Canberra, the disaster samples from each other. The laboratory will also need to document the number and the type of analysis performed on every sample that is tested. Custom Essay School. In table 1 there is an overview of the documentation that are generally noted on test samples . Table 3: Coding that should be present on test samples  Victim samples Personal effects samples Kinship samples. Identity of laboratory that performed extraction and analysis* Victim identification number Victim identification number. Extraction attempt number Identity of laboratory that performed extraction and analysis* Relationship to victim. Type of analysis Extraction attempt number.
Plate number, tube number, well number, etc. Type of analysis. Plate number, tube number, well number, etc. *These are usually noted in a multi-laboratory response. The recovery and preservation of samples are very important steps for the identification of human remains, especially when they are highly fragmented. It is very important for samples to be collected as soon as possible. But depending on text Justin-Siena High School the crash site and the environment it sometimes take much longer.
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With samples from a mass disaster it may not be possible to writing services Trafalgar Castle, still extract samples in that order. But DNA extraction from victim’s samples and reference samples should be separated by time and/or space. Help. DNA typing relies on custom writing Trafalgar the success of DNA extraction that yields enough quantity, quality and themes for essay writing School, purity. Because of essay writing services School, this naturally the critical is SKODA AUTO most desired extraction methods are the ones that minimizes DNA loss and the ones that overcome, removes or dilutes enzymatic inhibitors . The DNA IQ’ System (Promega Corp, Madison, WI) Current techniques that include organic solvents and custom writing services Trafalgar Castle School, ion exchange resins are time consuming, use more than one centrifugation step, uses toxic organic solutions and/or do not remove PCR (Polymerase Chain Reaction) inhibitors well enough. A system that was used for the extraction of themes for essay writing Hult International School, WTC samples proved to be the best. The DNA IQ’ System (Promega Corp, Madison, WI) uses a magnetic resin that the DNA can bind to. Essay School. It also has a denaturing agent that disrupts many types of cells/tissue.
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The lysing of the cell is carried out by using detergents Proteinase K and dithiothreitol (DTT) which are present in the lysis buffer. They insure that proteins and how to write (Navitas), cell debris are separated into organic phase and essay writing, the DNA remaining in the aqueous phase. They make sure that the cell membrane lyse, separate histones and denature histone proteins and critical AUTO University, destroy protein structures . Custom Essay Services. In the essay The University of Canberra third step the proteins and cell debris are removed by adding phenol chloroform isoamyl alcohol to the mixture. The phenol present wont mix with water and the proteins and debris will have an affinity with the organic phase. After centrifuging the mixture will have visible layers.
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A schematic overview of the chelex extraction is (Navitas), shown in figure 5. Polymerase Chain Reaction (PCR) After extraction is completed the DNA goes through PCR. PCR is an custom essay services Trafalgar enzymatic process in which a specific DNA fragment is being replicated over and over again. The process works by the heating and cooling the DNA extracted samples in a precise thermal cycle pattern for ?? 30 cycles . The PCR has three steps that forms one cycle . In each cycle it starts with denaturing the DNA template by a essay Tilton School heat. In the second step the School temperature is cooled so that primers can anneal (bind) to the DNA. In the last step the themes for essay Hult International Business School temperature is raised again at custom essay writing Trafalgar Castle the optimal temperature for DNA polymerase so it can activate. Text High. The DNA polymerase extends the template strands (see figure 6) .
When the cycles are repeated multiple times, you will get an exponential accumulation of your desired DNA fragment when present. Even with a small amount of isolated DNA, the PCR can generate a large amount of DNA . The primer, which is mentioned above, is the most important PCR component. The primer is a short DNA sequence that positions itself before the custom essay writing services Castle DNA fragment that needs to be copied. This means that the primer serves to identify the DNA fragment that needs to be copied. It is added in high concentration in relative to the DNA template in order to drive the PCR reaction. Other components that are added is naturally your template DNA that is going to be copied. Also added to the mix is a four nucleotide building blocks called dNTP`s and DNA polymerase that helps put the building blocks in text High the correct order on the template DNA.
The DNA polymerase used is Taq polymerase, which comes from a bacterium called Thermus aquaticus that inhibits hot springs. When setting up multiple samples that use the same primers, a master mix is made. From the master mix an equal amount of volume is pipetted in the PCR tube. By making this master mix you are insured homogeneity between samples and the pipetting of small volumes can be avoided . In this version of the PCR more than one DNA fragment will be copied simultaneously. This is achieved by adding more than one primer to the mixture. Essay Writing Services School. In order for it to work properly, the help a essay Tilton School primer pairs added should have similar annealing temperatures. With the adding of extra primer pairs the complexity of primer interactions increases. Excessive regions of complementarity has to be avoided in order to prevent primer-dimer formation. Primer-dimers are when added primers bind to custom essay services Trafalgar School, each other instead of the template DNA. Critical Thinking Is SKODA University. The possibility of primer-dimer formation increases with every new primer pair that are added .
After DNA extraction, DNA quantification is essential for polymerase chain reaction (PCR) based analysis. This is custom essay writing Trafalgar Castle School, important because a low concentration is preferable when working with multi-plex PCR. Generally an amount of 0,5 to 2,0 of DNA is optimal when using STR kits. AUTO University. When a higher amount of DNA is used, this makes the interpretation of results harder and more time consuming. With a low amount of essay writing services Trafalgar Castle, DNA, this can result in loss of alleles. Writing A Essay. This makes it harder to properly compare profiles with each other . Real-time quantitative PCR (qPCR) Like it was indicated above, quantification test is custom essay writing School, performed to determine amount of DNA that can amplified. A test that can indicate the quality and quantity of extracted DNA or PCR products, is beneficial for determining steps that need to be taking for the analysis of the DNA.
The most common approach to writing Tilton School, perform a qPCR is to use a flourogenic 5? nuclease assay, better known as TaqMan or with the use of an intercalating dye, like the SYBR green, that is specific for double stranded DNA molecules. The TaqMan monitors changes of a fluorescence while the SYBR green detects formation of PCR product . Probes for the TaqMan has two fluorescent dyes that emit at custom essay writing services Trafalgar School different wavelengths. Essay Istituto London. The probe hybridizes specifically in custom writing services Castle School DNA target region between two PCR primers. Help Writing A Essay. The probe has a slightly higher annealing temperature the primers. This way hybridization of the probes can begin when extension of primers begin.
At the 5? end of the probe, a reporter (R) dye is attached, while at the 3? end a quencher (Q) dye is synthesized See figure 7. While the probes are still attached together, fluorescence won’t occur, because of the suppression due to energy transfer of the two. When polymerization starts during the PCR run, any TaqMan probe attached to target sequence will be displaced. Because Taq polymerase has a 5? exonucleae activity. This means that it will chew at the TaqMan probe attached. When the dyes are released from the probe, they will begin to Trafalgar, fluorescence. This means that, the more fluorescence that is signaled, the critical Justin-Siena more target sequence is present as complementary to the probe . Real-time PCR analysis. The PCR process is essay writing services Trafalgar, defined in three phases. The exponential phase, linear amplification and plateau region.
These regions can be made visible in a plot of fluorescence versus the PCR cycle they are on (see figure 8). There is a high degree of precision of the making of new PCR products during exponential amplification. At 100% efficiency, there should be a doubling of PCR products with every PCR cycle. In a plot of cycle numbers versus log scale DNA concentration, will have a result of a linear relationship during the exponential phase of the polymerase chain reaction (see figure 8). The next phase is the linear phase. In this phase concentration of help writing, components falls and amplification efficiency slows down. This happens in an arithmetic increase. The final phase is the plateau region. Custom Essay Services School. This is where the production of the PCR products slowly stops. Multiple components that are being used for the amplification have reached the end.
The signal that is essay, being emitted slows down, causing the plateau to level out. The exponential phase of the real-time PCR analysis is the optimal place for measuring. This is the phase where the PCR product and the input DNA amount are most likely to essay Trafalgar, be consistent. The instruments use for writing Cesar Сolleges, real-time PCR uses a cycle threshold (CT) its calculations. This value stands for the terms of essay writing Trafalgar School, cycles when the level of fluorescence passes a threshold that is set by the software of the critical thinking text Justin-Siena real-time PCR instrument. The less cycles that is used to get to this threshold, the higher the input DNA molecules that was present in PCR reaction. This is why there is writing Castle School, a plot of log DNA and for essay writing Hult School, CT for every sample in a linear relationship with a negative slope. A rise in fluorescence can be traced to initial DNA template amount, when it is custom services Trafalgar Castle School, compared to samples that has known concentration. In figure 8, there are five samples (a, b, c, d, e) that were used in order to develop a standard curve. If the samples used have a good consistency and writing a essay Tilton, precision, a sample that has an unknown concentration can be calculated from the standard curve . Short Tandem Repeat (STR) analysis.
STR analysis is the first method of analysis that is used . 99,9% of the DNA in humans are the same. In the 0,1% region of DNA short tandem repeats are present as the region that exhibits a large variation between individual . STR`s are short DNA sequence that keep repeating. For example in the DNA sequence ATTCGCATCATCATCATCATCATCATCGCCA, the sequence ATC is repeated 7 times. Short tandem repeats are present at the same position on chromosomes in writing Trafalgar Castle School the human genome, but the write essay Istituto repeat unit varies per individual. With the writing Trafalgar Castle help of PCR reaction a STR analysis can be performed.
Primers that can be labeled binds to the DNA at a specific STR loci and multiplied. Because of the label that is attached at the primer, the amplified products can be detected at the end of the PCR reaction. By analyzing more than multiple STR`s at once, a DNA profile that is unique enough to identify an individual can be made . Sometimes a DNA sample is critical Justin-Siena, too degraded to get a DNA profile using the standard STR markers. In order to still be able to use this degraded DNA to get a profile, primers were repositioned. By repositioning the primers closer to the repeat region that needs to be amplified, the custom essay services Castle amplified product that is resulted is writing, much smaller.
As a result it makes genetic characterization of the sample for more STR`s, other that is used traditionally. This makes STR miniplex invaluable for custom essay, degraded samples . Single Nucleotide Polymorphism (SNP) analysis. By using the SNP`s genetic markers, the PCR amplified product can be much smaller than is possible in a STR miniplex . SNP is a single nucleotide mutation that occurs in the DNA sequence and is also the most common polymorphism in the human genome. These mutations can be a result from DNA replication or chemical damage. Justin-Siena School. A polymorphism is a variation/mutation in the DNA sequence that has a frequency of essay Trafalgar Castle School, more than 1% in the population. SNP occurs once every 1000 base pairs and tends to remain stable in themes writing Hult International School the population . For example in a known DNA sequence of CCTAA, a mutation of the second C to a T occurs (see figure 9). The amplified PCR product that can result from using SNP markers can be reduced by custom essay services Trafalgar School up to 60-80 bp.
This makes severely degraded DNA samples that could not be typed using STR`s still typable. Using a method that was validated during the WTC identification effort, typing on samples that were much compromised was still able to be identified. The method uses a florescent detection system. By labeling the possible alleles with a fluorescent dye, the signal of the dyes could be compared . Mitochondrial DNA analysis. Mitochondria is an organelle that resides outside the a essay Tilton School cell nucleus.
They contain their own chromosomal DNA. mtDNA are smaller, circular in custom writing Trafalgar Castle shape and are inherited maternally. They consist of approximately 16,569 base and have a high copy number in a cell. Thinking. Because mtDNA is passed down maternally, it is writing School, a good method to perform when reference samples are taking from maternal relatives. Using mtDNA can be an advantage when dealing with very small or degraded extracted DNA. Because of the high copies number of mtDNA that is available, an identification can still be possible using mtDNA markers, instead of STR markers that are found in nuclear DNA . There are several differences in the characteristic of the mtDNA and the nuclear DNA. The first being the size of the themes writing Hult Business genome. Nuclear DNA has about 3,2 billion bp and is 99.75% of total DNA. Services Trafalgar. While mtDNA has 16,569bp and 0,25% content DNA per cell. Help Writing A Essay Tilton School. While mutation rata for nuclear DNA is low, mtDNA has a 5-10 times higher mutation rate. The sequence of the mitochondrial DNA is custom essay writing services, a functional one and also highly conserved, it’s because of this that there isn’t a very big variation between individuals.
But there is non-coding D-loop (1000bp long), also known as control loop, that has two hypervariable regions called HV1 and HV2 (see figure 8). The variations observed in this part of the DNA tend to themes writing Hult International School, be Single Nucleotide Polymorphisms (SNPs). The length of the mtDNA doesn’t change with the presence of the SNPs and that’s why this are the regions that are used for forensic analysis. Due to the lack of DNA reparation, mtDNA has a large mutation. This causes the variations between individuals. Custom Essay Writing Services Castle School. The variation is not a lot, with HV1 and HV2 having only critical is SKODA AUTO University, a 1-3% difference between individuals who are not related . When analysis is performed, mitochondrial DNA is extracted and then with the help of Polymerase Chain Reaction (PCR), the essay writing services Trafalgar School HV1 and HV2 regions are amplified. Next step is to determine the sequence of the amplified product by using DNA sequencing.
After this samples can be compared to each other to themes for essay International School, get a possible identification . The Y-chromosome is small and found in only male individuals. This chromosome is altered through infrequent happening of a mutation. The combination of alleles between father and son is theoretically the same, if mutation does not occur . The advantages of custom writing Trafalgar Castle, Y-chromosome analysis are that in a mixture profile with female DNA for critical thinking Justin-Siena High, example in a sexual assault case, male specific amplification can take place to separate the two. Custom Essay Services Castle. Mixtures in fingernail scraping and saliva on skin can also be analyzed. Another advantage is critical thinking AUTO University, that it facilitates tracing family lineages in the paternal transmission. Limitations that arises are that it’s hard to distinguish between male family members when Y-STR typing is performed. Castle School. Duplications and deletions that are present in the DNA sequence, can make analysis more complicated. The Y-chromosome analysis is performed with the analysis of the STRs or SNPs. Write Essay Istituto. Y-STRs changes more rapidly and thus has more variables.
This makes it useful for forensic applications . Last step in analysis of writing, DNA is the separation and detection of the amplified DNA. In order to how to write a successful The University of Canberra, separate and detect them a capillary electrophoresis is used. A basic capillary electrophoresis apparatus is consists of a narrow glass capillary, two buffer vials, two electrodes connected to high voltage power supply, laser excitation source, fluorescent detector, autosampler for samples and a computer that controls the sample injection and detection. The capillaries are made out of glass, have an internal diameter of 50 ??m and is 50 cm long. The capillary which is a glass tube is filled with a polymer that serves as a gel in which molecules can migrate. Essay Writing Services Trafalgar Castle School. Samples are injected in the capillary when high voltage is applied, which helps to separate the DNA fragments. Fluorescent dye labels that are present in the DNA mixture are analyzed when they pass by a detector that uses a laser beam. The results are then analyzed and writing a good Ritz Сolleges, stored on a computer . A schematic of the capillary electrophoresis is shown in figure 9. DNA separation occurs when DNA fragments separate through the capillary when voltage is applied.
Smaller fragments go through the custom writing services Trafalgar capillary faster than the bigger fragments. The time is converted to base pair size with the help of internal size standards. Several components can affect the DNA separation. The polymer used, the capillary, the buffer and the voltage used . The two primary polymer are POP-4 and POP-6. The POP stands for performance optimized polymer and is made out of a linear, uncross-linked dimethyl polyacrylamide. The 4 and 6 in the POP, stands for the 4% and 6% concentration of this component. The POP-4 is used for the analysis of STR typing and POP-6 for DNA sequencing. Recently a Pop-7 has been introduced that can be used for both the for essay writing International Business School STR typing and DNA sequencing . The buffer is used to stabilize and solubilize the DNA. Writing Castle School. It also provide a charge for essay, the electrophoresis current.
Buffer concentration is an important aspect, because when it’s too high this can result in overheating of the capillary and custom, a loss of resolution. In order to avoid any problems with the running of the write Istituto electrophoresis, the buffer should be replaced regularly with a fresh sample . The capillary is central for the separation during the electrophoresis. Services Castle. The capillary is made of glass or fused silica. The inner walls also have a hydroxyl groups that are negatively charged. A Good Essay. Positive ions from the buffer, creates a double layer.
The movement of ions creates an custom School electroosmotic flow, that can be reduced or eliminated when a coating is added to the inside layer of the thinking High School capillary (see figure 10). Custom Essay Services Trafalgar Castle School. Uncoated walls create problems for critical University, reproducible DNA separation. Suggested is to change the capillary after 100-150 runs or whenever a decline of essay writing services Trafalgar, resolution is noted . Sanger method for DNA sequencing is a process where the thinking High use of polymerase incorporation is involved. The polymerase incorporation uses dideoxyribonucleatide triphosphates (ddNTPs) as chain terminators that is in turn followed by a separation step that is capable of single nucleotide resolution. Because there is no hydroxyl group at the end of the 3? ‘end of the nucleotide with a ddNTP the growing chain cannot continue growing. In the essay writing services Trafalgar Castle School mix there are both dNTPs and ddNTPS present, this way some DNA portions are still able to continue growing. Therefor at the end of the reaction there will be a series of strands that only differs one molecule from Tilton, each other.
See figure 11 for a schematic overview of the DNA sequence. Each DNA strand uses one forwards or reverse primer in a separate reaction. Attached to the ddNTPs are fluorescent dyes. ddTTP (thymine) has a red dye attached to it, ddCTP (cytosine) a blue label, ddATP (adenine) a green dye and lastly, ddGTP (guanine) a yellow dye. After the custom writing services Trafalgar School reaction is over, the sample goes through capillary electrophoresis, which separates the strands. In the results of the electrophoresis, the sequence of the DNA strand is shown . DNA Databases and CODIS. A DNA database is a database which stores DNA data and is used for analysis. There are different types of help a essay Tilton, DNA databases.
The National DNA databases are maintained by the government for storing DNA profiles of custom essay writing services Castle School, its population. These DNA profiles are based on PCR and STR analysis. DNA databases serve as valuable tools in aiding investigations. Write Istituto London. CODIS (COmbined DNA Index System) is a DNA database that consists of custom writing Castle, 13 STR loci’s. These are CSF1PO, FGA, TH01, TPOX, VWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51 and D21S11. Also the critical thinking AUTO Amelogenin is used for gender determination   . To make the job easier and Castle, avoid confusion with buffers it is better to how to write essay The University, use a complete STR kit. These kits come with their own buffer and control DNA. There are a lot of distributors of complete STR kits. Custom Writing Services School. It all depends on the research what kind of kit there is needed. Multiplex STR kits allows the amplification of the 13 CODIS loci .
The most widely known and used kits are from Promega. London. The product components normally consists of a buffer, primer pair mix (again, it depends on the research what kind of custom essay writing Trafalgar Castle, primer that will be used), control DNA. These are for how to a successful essay The University of Canberra, the PCR product. After the PCR, the custom writing services School kits come with an Allelic Ladder mix and an internal standard to help writing Tilton, be used for the gel. Chemicals that sometimes should be supplied that do not come with the kit are Nuclease-free water and taq DNA polymerase. The kits come with their own protocol and should be used in custom essay writing Castle School that specific order . Here is an example of the PowerPlex 16 HS System, which allows co-amplification of the 13 CODIS loci and Amelogenin for gender determination. All loci are amplified in a single tube and analyzed in a single injection and is also compatible with ABI. This system consist of Powerplex HS 5X Master Mix, Powerplex 16 HS 10X Primer Pair Mix, 2800M Control DNA and water (Amplification Grade) in the Pre-amplification components box. In the Post-amplification components box there is a PowerPlex 16 HS Allelic Ladder Mix and Internal Lane Standard.
How the mixes for the PCR amplification should be made is displayed below in help writing a essay Tilton figure 10  . Statistical interpretation of DNA profile. When it comes to getting a match with a DNA profile, it’s important to determine if the DNA profile are from the same individual or if maybe there is another individual with the same profile. It’s practically impossible to get a DNA from every individual. In order to still determine a match probability, Allele frequencies from different ethnic groups, from services Castle, validated databases are collected . More than one data point will need to be collected in order to get a reliable estimation of allele frequency.
There has to be a minimum where the assurance is there that an allele has been sufficiently sampled for use in statistical test. An allele has to be observed for critical AUTO University, a minimum of five times in order to Trafalgar Castle School, be qualified for use in a statistical calculations. The frequency for the minimum allele is thinking AUTO University, determined using the formula 5/ (2N). N stands for the number of services Trafalgar, individuals and 2N for chromosomes counted before they pair . In order to calculate the frequency of critical thinking text Justin-Siena, a DNA profile, the frequency of every allele analyzed are multiplied. The frequency of the writing Castle School allele can differ dependent on the population database.
The database that is used needs to be sufficiently large and representative of the population of the individual whose DNA is being examined . In figure 12, DNA profile frequencies are given for Tilton, the U.S. Caucasian. Take for example allele 11 of D13S317 shown in figure 12. Custom Writing Services. Samples of 604 alleles were taken where it was observed 205 times. This translates to an allele frequency of 0,33940. In other words, there is a 34% chance that a random individual will have the same allele. The same goes for allele 14 of D13S317. Writing Hult Business School. This allele was observed 19 times in a population of 604. Frequency is therefore q=0,04801. Writing Trafalgar. So an individual with the alleles 11 and 14 for the D13S317, can be calculated using the formula 2pq (p standing for the first allele and q standing for thinking is SKODA University, the second allele).
Putting the custom frequency calculated in the formula creates 2 x 0,33940 x 0,0481= 0,0326. This frequency indicates that ??3% of the U.S. Caucasian population. The frequency of all the genotypes will then be multiplied to get the for essay International frequency of the DNA profile. In case of the DNA profile in figure 12, this comes to a combined frequency of 8,37 x 1014. Services Castle School. So 1 in 837 trillion . In order to ensure that DNA testing is being performed correctly there are standards that the laboratories has to adhere to. These standards has to assurance the quality of the work.
Every laboratory should have a quality assurance program in place. Organization and management. First thing is to AUTO University, have a management staff that the authority and means to carry out the duty that is Castle, assigned to them and keep the quality assurance in the laboratory. Help Writing. There should also be a technical leader that is in charge of the essay Trafalgar technical operations. If the CODIS is in use at the laboratory, there should be a casework CODIS administrator who is accountable for CODIS on site.
The laboratory should also have two full time employees who are qualified DNA analysts. The responsibility, authority and interrelation of every personal operational at the laboratory has to write Istituto Marangoni, be specified and documented. Custom Castle. Lastly a contingency plan has to be in planned and approved in the case that a technical leader position is vacated . The employees of the laboratory has to have the proper education and writing Ritz, experience. The job description of the personnel needs to writing services Trafalgar Castle School, have a written job description that can be expended with documentation of the responsibilities, duties and skills. The laboratory needs to be in write Istituto Marangoni possession of the training program for qualifying all analyst. Included are all the custom essay writing services DNA analytical procedures that will be performed by the personnel. Also all the practical exercises needs to include the examination of a range of samples that are routinely analyzed at the laboratory. The training program needs to teach and write Istituto Marangoni London, asses the skills and ability to perform a successful DNA analysis. The trainee needs to give an individual demonstration of capability, which shall be documented by essay Trafalgar Castle the laboratory. When a new analyst is hired the technical leader is tasked with accessing the writing persuasive essay Ritz Сolleges new hire`s previous training and be responsible for the proper documentation.
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Attendance needs to be formally documented and when the custom writing services Trafalgar School program is finished the technical leader reviews and approves it. Critical Text School. The laboratory also needs to have an approved program that is used for annual review of writing Castle, scientific literature. This way the analyst ongoing reading of scientific literature is documented. The laboratory needs to have access to help writing a essay Tilton, collections of information’s containing DNA analysis . The technical leader also has certain qualification that needs to be tended to. The minimum education requirements are a Master`s degree in biology, chemistry or forensic science.
In addition they also need to have a successfully completed 12 semester or equivalent credit hours in the subject’s biochemistry, genetics, molecular biology and statistics or population genetics. In the custom essay services Trafalgar Castle 12 semester completed, there needs to be at least one graduate level course for the period of 3 semesters. The technical leader of also needs to have a minimum of three years of critical AUTO, experience in a forensic DNA laboratory, in which the laboratory participated in the identification and evaluation of biological evidence in writing services Trafalgar Castle School a criminal matter capacity. The duties of the technical leader includes overseeing the technical operation of the laboratory and being able to make decision of initiating, suspending od resuming DNA analytical operations. Other responsibilities include the evaluating and documenting of the themes writing International validation of methods used and proposing new or modified analytical procedures.
Also reviewing academic documents, training records and approve qualifications of essay School, new analyst before they are allowed to perform analysis on casework independently. The technical leader has to approve specifications for outsourcing agreements and review internal and external DNA audit that might needs to be corrected and documented. Help Writing A Essay School. Lastly the technical leader is custom essay writing Trafalgar School, responsible for annually reviewing and documenting of laboratory procedures and reviewing and approving of training, quality assurance and proficiency testing. The technical leader needs to be a full time employee and needs to write London, be accessible for consultation onsite, via the telephone or electronically . As an employee of the Trafalgar laboratory, the analyst needs to have a minimum education of a good Cesar, a bachelor`s degree or advanced degree in biology, chemistry or forensic science related area, The analyst is expected to have completed the course work successfully with subject areas of biochemistry, genetics, molecular biology, statistics and population genetics. The analyst needs to have at least six months of experience in forensic human DNA laboratory experience. Essay Services Castle School. The experience of the analyst needs to be documented and if needed, experience augmented by additional training of DNA identification handlings of the laboratory. Before participating in casework, the analyst needs to complete analysis of a range of samples that the laboratory regularly encounters and complete a competency test . Laboratories needs to be designed in a manner where the integrity of analyses and evidence is ensured. Entry to the laboratory needs to be controlled in order to themes for essay writing International Business School, prevent unauthorized personnel.
When keys or combinations are handed out, these needs to be documented and limited to custom essay writing, personnel that is designated by help writing School laboratory management . Before PCR is custom services Castle, performed, the DNA extraction and persuasive essay Cesar, PCR set up, needs to be performed in either separate times or separate space. The amplified DNA, needs to be developed, processed and maintained on a room(s) that is separate from where the custom essay services Trafalgar evidence examination, DNA extraction or PCR setup areas are performed. Thinking Text Justin-Siena High. A robotic workstation can be used, only custom essay services Trafalgar Castle, if the writing a good persuasive analytical process has been validated. In that case DNA extraction, PCR setup and amplification can be performed in custom Trafalgar Castle School a single room. Writing A Essay School. Lastly procedures for the cleaning and essay writing Castle School, decontaminating of the write a successful essay The University facilities and its equipment needs be in place and followed . In order to ensure the integrity of the physical evidence, the laboratory needs to have a documented evidence control system.
Evidence needs to custom Trafalgar Castle School, be marked with a unique tag on package of the evidence. A definition of the text Justin-Siena High evidence and work product needs to be clearly defined. When a tag is not used, a system for distinguish samples when they are being processed needs to be devised and put in place. Custom Essay Writing. Chain of custody needs to be documented and maintained, either on persuasive essay Cesar Сolleges a hard copy or in an electronic format. A signature or something equivalent in case of an electronic format needs to be present for all who received and transferred the evidence, with corresponding date. Documented procedures needs to be in essay Castle place in order to minimize loss, contamination and deleterious change of evidence.
A proper placement for evidence storage and work product that is still in progress needs to be in place, which is controlled and secure . Analytical procedure that are to write The University, be had and followed, needs to be approved by essay writing services Castle School the technical leader. A Good Persuasive Сolleges. The procedures needs to be reviewed annually. Every analytical method needs to have their own standard operating procedure that needs to specify items like the reagents, sample preparation, extraction methods, equipment and controls. A written procedure for the documentation of commercial and formulated in-house reagents needs to be in place. Label commercial reagents with the name of the custom services Trafalgar reagent and expiration date. Formulated in-house reagents are to be labeled with the name of reagent, either the date of preparation or expiration date and the analyst who formulated the reagent. Critical reagents in test kit or systems for quantitative PCR and genetic typing needs to be identified and evaluated before they are used on evidence samples. Thermostable DNA polymerase, primer sets and write Marangoni London, allelic ladders, also needs to be identified and writing Trafalgar School, evaluated .
Before DNA amplification, evidence samples needs to be quantified. Only when a validated method system is used that have shown to give reproducibly and reliably yield a successful DNA amplification, quantification is not needed. When quantifying, quantification standards are to writing, be used. The positive and negative control needs to be amplified at the same time as the case samples at the same loci with the same set of primers as the case samples. Blank reagent controls that are being used for extraction, are to be extracted simultaneously, amplified by using the same primers, instrument and conditions as the case samples and typed by using the same instrument and injection conditions. The DNA procedures used by the laboratory are to custom essay writing Trafalgar Castle, be checked annually or when a major change is made in the method . Procedure for the taking and maintaining casework notes needs to be in place. These needs need to support conclusion taken in the reports. The laboratory needs to maintain and retain, in hard or electronic form, all notes and documentation from analyst regarding their analysis. The casework reports needs to have the case identifier, a description of the critical thinking is SKODA evidence that was examined, description of the method used, the amplification system, results and/or conclusion, an interpretive statement, date that report is issued, the disposition of the evidence and lastly a signature and title . Twice a year, with the first one occurring in the first six months and the second in the last 6 months f a year, a proficiency testing needs to be undergone by custom essay writing Trafalgar Castle School the analyst, technical reviewers, technicians and personnel appointed by the technical leader.
Workers who regularly use manual and automated methods, will need to School, go through a proficiency testing at least once a year. New workers needs to go through proficiency testing in the first six months. The proficiency testing needs to be defined, documented and custom essay Castle School, use the correct dates. The exact items from the proficiency testing that needs to be recorded are the test set identifier, the analyst/participant, date of analysis and its completion, copies of data and notes that supports the conclusions taken, the proficiency test results, discrepancies and if needed the corrective actions that were taken . 1. Lessons Learned From 9/11: DNA Identification in Mass Fatality Incidents https://www.ncjrs.gov/pdffiles1/nij/214781.pdf. 2. Cesar. M.prinz, A. Carracacedo; DNA Commission of the International Society for Forensic Genetics (ISFG): Recommendations regarding the role of services Trafalgar, forensic genetics for disaster victim identification (DVI); Forensic Science International: Genetics; Pages 3-12; 2007. 3. How To Essay The University Of Canberra. B. Budowle, F.R. Bieber, A.J.
Eisenberg; Forensic aspects of mass disasters: Strategic considerations for DNA-based human identification; Legal Medicine; page 230-243;2005. 4. Mo Bi Tec molecular biotechnology. 5. NFSTC Science serving justice. 6. Forensic DNA typing STR: Page 62-64, 84-86,92,93. 7. Butler John M.; Advanced topic in forensic DNA typing: methodology; 2012; Elsevier, Maryland USA, page 371, 372, 141-150,417,418. 8. Sequencing, forensic analysis and genetic analysis http://www.atdbio.com/content/20/Sequencing-forensic-analysis-and-genetic-analysis#Short-tandem-repeats.
9. Sirius genomics. 10. Forensic science central; DNA. 11. FBI biometric analysis. 12. Promega technical manual for the powerplex 16 system. 13.
Butler John M.; Fundamentals of services Castle School, forensic DNA typing; 2010; Elsevier; Maryland USA; page 111-121, 229, 238-241, 251, 252. 14. Quality Assurance standards for forensic DNA testing laboratories. DNA analysis workload estimated worksheet. Search our thousands of essays:
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