Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin

Alzheimer is one of the diseases that destroy brain cells, causing problems with thinking, memory and behavior. Patients' quality of life is compromised because the disease can affect work, social life and lifestyle. So, it is vital to determine and identify this disease at early stage so that...

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Main Author:	Sha-aladin, Shaira Shuwari
Format:	Thesis
Language:	English
Published:	2008
Subjects:	Pharmacopoeias Pharmaceutical chemistry
Online Access:	https://ir.uitm.edu.my/id/eprint/101233/1/101233.PDF
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id	my-uitm-ir.101233
record_format	uketd_dc
spelling	my-uitm-ir.1012332024-09-28T23:39:35Z Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin 2008 Sha-aladin, Shaira Shuwari Pharmacopoeias Pharmaceutical chemistry Alzheimer is one of the diseases that destroy brain cells, causing problems with thinking, memory and behavior. Patients' quality of life is compromised because the disease can affect work, social life and lifestyle. So, it is vital to determine and identify this disease at early stage so that later modification can me made as it also can be caused by environment factor besides genetic factor. Apolipoprotein E-£4 allele was found to be the major genetic risk for Alzheimer's disease. Therefore, this study is aimed to develop Allele specific - polymerase chain reaction (AS-PCR) method, validate the method and later use it for detecting of the types and frequencies of the genetic variants of APOE. After DNA extraction, the DNA was used as samples in PCR reaction to determine the APOE polymorphism by using allele-specific primers. We have found the most suitable primers (FI wt, FI mt, RI cm, F2wt, F2mt, and R2cm) for multiplex PCR to detect the polymorphism of APOE genotype. Temperature of 54 °C was found to be the most suitable for these allele-specific primers to anneal to their specific complementary sequences in the template DNA. Touchdown PCR was shown to increase specificity of PCR reaction in this study. Addition of 5% DMSO also improved the specificity and PCR yield. Optimisation of PCR was shown to be successful either by increasing the sensitivity or the specificity of PCR reaction. Increase PCR sensitivity means that increase detection of true target sequence while improving the specificity of reaction result in amplification of the correct sequence. 2008 Thesis https://ir.uitm.edu.my/id/eprint/101233/ https://ir.uitm.edu.my/id/eprint/101233/1/101233.PDF text en public degree Universiti Teknologi MARA (Kampus Puncak Alam) Faculty of Pharmacy Darawi, Mohd Nazif
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Darawi, Mohd Nazif
topic	Pharmacopoeias Pharmaceutical chemistry
spellingShingle	Pharmacopoeias Pharmaceutical chemistry Sha-aladin, Shaira Shuwari Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin
description	Alzheimer is one of the diseases that destroy brain cells, causing problems with thinking, memory and behavior. Patients' quality of life is compromised because the disease can affect work, social life and lifestyle. So, it is vital to determine and identify this disease at early stage so that later modification can me made as it also can be caused by environment factor besides genetic factor. Apolipoprotein E-£4 allele was found to be the major genetic risk for Alzheimer's disease. Therefore, this study is aimed to develop Allele specific - polymerase chain reaction (AS-PCR) method, validate the method and later use it for detecting of the types and frequencies of the genetic variants of APOE. After DNA extraction, the DNA was used as samples in PCR reaction to determine the APOE polymorphism by using allele-specific primers. We have found the most suitable primers (FI wt, FI mt, RI cm, F2wt, F2mt, and R2cm) for multiplex PCR to detect the polymorphism of APOE genotype. Temperature of 54 °C was found to be the most suitable for these allele-specific primers to anneal to their specific complementary sequences in the template DNA. Touchdown PCR was shown to increase specificity of PCR reaction in this study. Addition of 5% DMSO also improved the specificity and PCR yield. Optimisation of PCR was shown to be successful either by increasing the sensitivity or the specificity of PCR reaction. Increase PCR sensitivity means that increase detection of true target sequence while improving the specificity of reaction result in amplification of the correct sequence.
format	Thesis
qualification_level	Bachelor degree
author	Sha-aladin, Shaira Shuwari
author_facet	Sha-aladin, Shaira Shuwari
author_sort	Sha-aladin, Shaira Shuwari
title	Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin
title_short	Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin
title_full	Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin
title_fullStr	Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin
title_full_unstemmed	Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin
title_sort	development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein e / shaira shuwari sha-aladin
granting_institution	Universiti Teknologi MARA (Kampus Puncak Alam)
granting_department	Faculty of Pharmacy
publishDate	2008
url	https://ir.uitm.edu.my/id/eprint/101233/1/101233.PDF
_version_	1811769151244468224

Development of allele specific-polymerase chain reaction for detecting genetic polymorphism of apolipoprotein E / Shaira Shuwari Sha-aladin

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