PCR Amplification-Marfan Syndrome (Lab Report Sample)
The lab report should be (800-2000 words) excluding table of contents and references.
1. Page size: A4
2. Page margin: Left margin 1.5 and all other sides 1.0
3. Font type: Times New Roman
4. Font size:
For title: 16
For sub-headings: 14
For text: 12
5. Line spacing: 1.5
• Heading and sub-heading should be numbered.
• All pages should be numbered consecutively from the start till the end of the document.
C. Tables and Figures:
Tables or figures must be numbered and include legends. References for images and tables should be provided (these will not be included in the final word count).
Be aware that the Scientific Research Committee at the Faculty of Applied Medical Sciences approved 20% similarity as the acceptable percentage for plagiarism using the IThenticate software (easy level). It is the responsibility of the students to check their research report by the plagiarism detection software (IThenticate) to determine the level of similarity of the material they have used. The plagiarism check should include the main text only, without table of contents, methodology (if applicable), and references.
Please ensure that the plagiarism detected is not more than 20% plagiarism (copying word-for-word). If the plagiarism level is calculated to be above 20%, then 5% of the final assignment marks will be deducted for every 10% above this acceptable percentage.
If the percentage detected is more that 50%, the student needs to re-submit after paraphrasing to reduce this percentage. In addition, 5% of the final assignment mark will be deducted for every 10% above the acceptable percentage.
1. Use original scientific research article papers and do not use commercial websites (such as Google).
2. Include the references used within the text (as numbers in the bracket ( ), in the order it is appearing (Vancouver style) and full references (in the reference list) at the end of the assignment (not included in the word count or number of pages).
F. Arrangement of the assignment for submission:
1. Cover Page (Must be fully completed)
2. Table of contents with page number (start numbering from this page as page 1)
3. Lab report main text (Topic under different headings)
5. Print on one side of A4 size paper.
PCR Amplification-Marfan Syndrome
Course Number and Name
Contents TOC \o "1-3" \h \z \u 1.0 Background PAGEREF _Toc68172814 \h 22.0 Principle of the assay PAGEREF _Toc68172815 \h 23.0 Summary of the assay PAGEREF _Toc68172816 \h 24.0 Clinical Significance PAGEREF _Toc68172817 \h 25.0 Strategy for analysis PAGEREF _Toc68172818 \h 26.0 Specimen collection or acquisition PAGEREF _Toc68172819 \h 37.0 Criteria for rejected samples PAGEREF _Toc68172820 \h 38.0 Equipment and supplies PAGEREF _Toc68172821 \h 38.1 PCR Machines PAGEREF _Toc68172822 \h 38.2 Gel electrophoresis Apparatus PAGEREF _Toc68172823 \h 39.0 Reagents PAGEREF _Toc68172824 \h 310.0 Assay procedure PAGEREF _Toc68172825 \h 411.0 Analysis of results PAGEREF _Toc68172826 \h 412.0 Turnaround time and schedule for retaining the specimen PAGEREF _Toc68172827 \h 513.0 References PAGEREF _Toc68172828 \h 6
Marfan syndrome (MFS) refers to connective tissue disorders instigated by a genetic defect that could be hereditary. Connectivity tissues are found all over the body of an organism with multiple organs (1). Marfan syndrome is gender insensitive and can affect males and females in equal proportions, and its mutation isn’t biased on race, ethnicity or geographical location. The basic characteristic of people with Marfan syndrome is that they tend to be slender and tall with elongated legs, fingers, toes and arms. MFS is often caused by fibrillin-1 (FBN1), which is responsible for abnormal connective tissue. Almost all the patients with Marfan syndrome inherited it from their linage (1).
2.0 Principle of the assay
Marfan syndrome detection is more dependent on a sensitive procedure of detecting FBN1 mutation. Fibrillin-1 genes denoted by FBN1 are proteins located in the chromosomes, which cause Marfan syndrome. A restriction enzyme is used to differentiate genes by identifying a single base change in DNA. Genotyped DNA sample is digested using restriction enzymes to generate fragments separated using differentiations in their sizes through gel electrophoresis.
3.0 Summary of the assay
DNA samples extracted from patients' suspects are subjected to PCR using a primer set. After that, gel electrophoresis was used to detect FBN1 proteins from the PCR process products with restriction enzymes' help. The properties of the restriction enzyme used in this study are;
4.0 Clinical Significance
Assay test done on clinical samples through polymerase chain reaction (PCR) on patients’ reports rapid detection of Marfan syndrome (2). Various techniques can enhance the efficiency and specificity of test outcomes on patients suspected to be suffering from MFS. Although there is no absolute cure for Marfan syndrome, treatment is administered on case to case basis.
5.0 Strategy for analysis
The chronology of testing steps of MFS on samples collected from patients are; 1) extraction of DNA templates from the blood samples collected, 2) PCR amplification on the DNA sample extracted was amplified to obtain a single cell, 3) digestion is the process in which DNA is cut into specific sites, 4) gel electrophoresis is the step after digestion where the products of digestion are separated using the length of the molecule.
6.0 Specimen collection or acquisition
The blood specimen is ideal for assay testing of MFS through PCR and gel electrophoresis. The blood sample was taken from a 14-year-old girl in a 5ml sterile tube and was labelled with a unique identification code, the patient's name, and the collection date. The specimen was collected in the laboratory, and therefore, no shipment was required; nevertheless, it was maintained at an ideal temperature of 4-8OC.
7.0 Criteria for rejected samples
the specimen could be rejected if found that; 1) the equipment used were contaminated, 2) the specimen could have been exposed to extreme temperature, 3) when the label is lost or not legible, 4) In case the blood sample mixes with other substances in the laboratory.
8.0 Equipment and supplies
8.1 PCR Machines
A PCR machine is also known as a thermal cycler machine, and its main application is to amplify DNA by regulating temperature cyclically.
8.2 Gel electrophoresis Apparatus
Gel electrophoresis device splits charged molecules. The split occurs where differently charged molecules in terms of strength are put in an aqueous solution. They migrated at different speeds from different zones; thus, they are easily identified.
The following reagents were used in the PCR process of the DNA samples collected; 1) dATP-Deoxyadenosine triphosphate 2) dTTP-Deoxythymidine triphosphate. MgCl2 solution was also added to ensure that the enzyme works optimally.
10.0 Assay procedure
A blood specimen was taken from a 14-year-old girl who had visited a physician for a checkup on cardinal vascular-related ailments. She was suspected to be suffering from Marfan syndrome. The general considerations in preparation and processing of DNA extraction, testing and amplification are; always to put on gloves, use aerosol-resistant filter tips, using screwcap tubes, prepare reactions in replicate -ideally as triplicates. Use calibrated pipets dedicated to PCR, using a no-template control to ascertain no contamination in the specimen and apparatus.
The blood sample collected was screened for contamination and clotting to assess its suitability for the next procedure. DNA was then extracted from the blood specimen collection to be used as the template. The PCR amplification was conducted under standard conditions; a microtiter plate was used to hold the DNA set to be processed in a PMC3 thermocycler for PCR amplification (1). The followi
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