Additional Tests
The clinician requested HNF1A and HNF4A gene mutations genetic testing once analysing previous results. Before any further analysis could go ahead, the patient had to give consent for the genetic analysis having been briefed on the tests being performed and possible consequences.
Once consent was given, blood samples were taken in Craigavon Area Hospital by a registered nurse via venepuncture, using a tourniquet and an 18 gauge butterfly needle, collected into sterile purple topped blood tubes with added EDTA anticoagulant. A minimum of 10ml of blood was required, so three EDTA tubes were filled. In an attempt to monitor pre-analytic variables, the patient would have been advised to relax their arm while the bloods were
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The requester had to ensure they completed the form electronically and to send a printed copy along with the sample, ensuring the details on both the samples and the two request form were the same. (Example of the Genetic Test Request form is included in Appendix 1.) Once the electronic form was sent, the paper copy and the blood samples were sent to Craigavon specimen reception in the laboratories via porter where they were sorted, processed and checked before transfer to the Royal Devon and Exeter Hospital …show more content…
Sanger sequencing methodologies used for the HNF1A and HNF4A gene analysis is referred to as ‘first’ generation DNA sequencing and allows for the clinical determination of the order in which the bases appear in a given strand of DNA known as a gene.
Test methodologies for analysis of the HNF1A gene consisted of analysis of all the coding regions and the exon-intron boundaries regarding that specific gene. This was carried out by a DNA sequencing method known as Sanger sequencing Exeter laboratories on ABI PRISM 377 DNA Sequencer analyser. Likewise, the investigations carried out on the HNF4A gene was also carried out by the Sanger sequencing method. This time carrying out investigations of the P2 promoter region, as well as associated exon-intron boundaries of the HNF4A gene.
The ABI PRISM 377 DNA Sequencer is an automated instrument designed primarily for the analysis of fluorescently-labelled DNA fragments separated by gel electrophoresis. Gel electrophoresis is the method used to separate the DNA based on their size as the charged molecules migrate through a polyacrylamide gel when an electric current is
Figure 2 shows the results of the electrophoresis. Lanes 5 and 7 indicate the fragments obtained when the plasmids are digested with both restriction enzymes, indicating the approximate fragment size for the hlyA gene, the pK184 plasmid and the pBluescript plasmid. This is useful for identifying the recombinant DNA needed for this experiment
Amplification reaction was done in a 25.0 µL reaction mixture containing 0.4 µL DNA (from DNA extraction), 5.0 µL of 10X PCR reaction buffer, 14.2 µL of sterelized dH2O, 2.0 µL of magnesium chloride (MgCl2, 25 mM), 1.0 µL nucleotide/dNTP mix (10 Mm), and 0.4 µL of 5 u/µL Taq DNA polymerase for each primer namely respectively. The components and the volume used for the amplification reactions are listed in Table 3.2. For the reaction, PCR reaction was performed in a programmable gradient-enabled thermocycler (Bio-Rad MyCycler™ Thermal Cycler).
imported into an aparatus using gel electrophoresis to compare the sample of DNA to other
The given DNA ladder sample and each individual ligation samples were run on 40ml of 0.8% agarose in 1x TAE buffer for approximately sixty minutes at 110V. The appropriate volume of 6x GelRed track dye was used after it was diluted to a final concentration of 1x and incubated for thirty minutes. Finally, the gel was illuminated under UV light and analyzed.
Burlington Northern Santa Fe Railway was asked by the U.S. Equal Employment Opportunity Commission (EEOC) to “stop testing its employees for genetic defects as part of a workplace discrimination settlement”. (Schafer, 2001) This comes as a result of over 30 employees being tested without their knowledge or consent that genetic testing was being conducted using some of their blood samples provided during their physicals. This is the first case initiated against a company and a groundbreaking one when it comes to genetic testing.
Harmening, D. M. (2005). Modern Blood Banking & Transfusion Practices. Philidelphia, PA: F.A. Davis Company.
In Gattaca, the plot focuses on the ethics, the risks, and the emotional impact of genetic testing in the nearby future. The film was released in the 90s; yet in the present, the film does not give the impression of science fiction. Today, genetic testing is prevalent in many aspects of the scientific community. This paper will describe genetic testing, its purpose, diagnostic techniques that use genetic testing, relating Huntington’s disease to genetic testing, and the pros and cons of genetic testing.
By reading the DNA of a fetus Prenatal DNA sequencing is the next dividing line of the genome revolution. Prenatal DNA sequencing is a noninvasive screening in which scientist can be able to sequence the DNA before birth. This is a primary process of gene altering in which where researchers are trying to correct an imperfection in a child. Scientist has discovered by a mother giving her blood sample and the father giving a spit sample, an entire genome of their fetus can be born. Clinical test capitalize on this DNA to discern the baby’s sex and determine whether mother and child have incompatible Rh blood groups, which can lead to fatal complications. Every gene and every chromosome known about a baby’s genetic make-up, while still in the u...
Genomic sequencing is the process of determining the precise order of nucleotides within a DNA molecule. It includes any method or technology that is used to determine the order of the four DNA bases – thiamine, adenine, guanine, and cytosine– in the strand of DNA (NHGRI, 2011). In each organism, these bases are arranged in a unique and specific sequence, and it is this sequence that is the genetic code of the organism. Genomic sequencing has had an impact on nearly every field of biological research including human genetics and genomics, plants and agriculture, microbes, medicine, viruses and infectious diseases, environmental genetics and evolutionary biology. By first examining the development of gene sequencing technology we will be able to view its role in evolutionary biology, its contribution to phylogenetics, and how it has changed our understanding of the biological tree of life.
Modern techniques , rather than the gene map , maps the map of the DNA within the gene itself : the positions of short sequences " marker " are used as markers signaling over the cromosssomas . Once a gene is discovered, it is necessary to unravel its base sequence prior to its function being studied . The sequencing has become easier with the development of methods for cloning the DNA - producing large amounts of identical fragments. In the method most widely used DNA sequencing , the chain is denatured into single strands . These are then used as templates for DNA synthesis , but such that replication to as the double helix reaches a certain growth in the mold base . In addition to provide DNA polymerase and the four bases, A - G -C- T, also using small amounts of these dideoxynucleotide bases. This is incorporated , as the normal bases, the double helix growth but prevent the continuation of the chain. The fragments are then separated by gel electrophoresis and the base seq...
Testing of one kind or another has always been very common in the job application process, whether it’s the testing of one’s skill sets through an application or drug testing. Now, through scientific advances of the Human Genome Project, a new type of testing has become available to employers: genetic testing. This new sort of testing would detect any genetic problems in a potential employee’s DNA, including genetic predisposition for disease and possible health concerns in the future. Genetic testing could be beneficial to companies monetarily, as it would be a way to screen their employees for long term, damaging factors which could be expensive for the company. The legal right to do so, however, is a controversial issue in the business world. Many people regard it as an invasion of privacy, an open invitation for bias in hiring pools, and a violation of the Americans with Disabilities Act (French). Genetic testing could be used in both the hiring process, and as an attempt for companies to cater to their employees’ needs. It could also, however, be used as a tool of discrimination against those with minor or major genetic abnormalities. Many ethical issues are raised: does a person have a right to anonymity in their DNA? Should employers discriminate against those with genetic problems, a factor out of the subject’s control? These disputes and more are what make genetic testing in the workplace a contentious topic today. Unless an act, such as the Genetic Nondiscrimination in Health Insurance and Employment Act, is passed, discrimination will run rampant through the job market, injuring the modern employee in the process. Despite the quantity of information about employees that could be retriev...
"Testing Children for Genetic Status." Ama-assn.org. American Medical Association, 1995. Web. 2 Feb. 2014. .
The desire to have a "normal" child is held by every parent and only now are we beginning to have the ability to select for that child. In preparation to receiving genetic testing, the parents are required to meet with a genetic counselor. A detailed description of the testing methods are reviewed with the couple as well as the risks which are involved with each. Upon an understanding of the procedures, the counselor discusses the many possible outcomes which could be the result of the diagnosis. Finally, before any tests are performed, anxieties from either of the parents are addressed as well as the psychological well-being of the parents.
...to get his entire genome mapped out and was prescribed a medicine that help him to live fifteen more months. The risk of genetic testing is the patient finds out when they are going to die and it is nothing that he or she can do. Taking into account a person who finds out that they will have pancreatic cancer, most of the time people with pancreatic cancer die.
The scientific and medical progress of DNA as been emense, from involving the identification of our genes that trigger major diseases or the creation and manufacture of drugs to treat these diseases. DNA has many significant uses to society, health and culture of today. One important area of DNA research is that used for genetic and medical research. Our abi...