Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa

Mutations of human VKOR gene leading to warfarin resistance in certain patients and polymorphism which is associated with a significantly higher risk of aortic calcification in certain patients have been identified. By identification of VKOR gene defects, prevention or treatment inititative such as...

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Main Author: Mustafa, Nurul Ashikin
Format: Thesis
Language:English
Published: 2008
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Online Access:https://ir.uitm.edu.my/id/eprint/100891/1/100891.PDF
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spelling my-uitm-ir.1008912024-08-26T04:59:35Z Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa 2008 Mustafa, Nurul Ashikin QD Chemistry DNA. Deoxyribonucleic acids Mutations of human VKOR gene leading to warfarin resistance in certain patients and polymorphism which is associated with a significantly higher risk of aortic calcification in certain patients have been identified. By identification of VKOR gene defects, prevention or treatment inititative such as gene therapy may be initiated for affected patients. PCR is a technique to amplify specific DNA sequences in vitro. PCR is able to rapidly amplify a specific region of a single DNA molecule to yield sufficient quantities that can be cloned, sequenced, or analysed by sequences mapping . The aim of this study is to amplify human VKOR gene using PCR for use in cloning and expression. In this study, specific primers flanking the complete VKOR coding region was designed. The specificity of the primers were evaluated using Oligo Explorer 1.2 Software. The PCR was then performed to detect the VKOR gene. The PCR conditions were optimized by varying the PCR temperature and time. In order to prove the target PCR products were obtained after amplification, the PCR products were separated by gel electrophoresis and visualized under UV transilluminator. The band of interest which was 492 base pairs in size were obtained after human VKOR gene amplification was performed. The amplicon can then be used for cloning and expression of YKOR enzyme. However, more studies need to be performed for more specific amplification without non-specific co-amplification. 2008 Thesis https://ir.uitm.edu.my/id/eprint/100891/ https://ir.uitm.edu.my/id/eprint/100891/1/100891.PDF text en public degree Universiti Teknologi MARA (Kampus Puncak Alam) Faculty of Pharmacy Salleh, Mohd Zaki
institution Universiti Teknologi MARA
collection UiTM Institutional Repository
language English
advisor Salleh, Mohd Zaki
topic QD Chemistry
QD Chemistry
spellingShingle QD Chemistry
QD Chemistry
Mustafa, Nurul Ashikin
Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa
description Mutations of human VKOR gene leading to warfarin resistance in certain patients and polymorphism which is associated with a significantly higher risk of aortic calcification in certain patients have been identified. By identification of VKOR gene defects, prevention or treatment inititative such as gene therapy may be initiated for affected patients. PCR is a technique to amplify specific DNA sequences in vitro. PCR is able to rapidly amplify a specific region of a single DNA molecule to yield sufficient quantities that can be cloned, sequenced, or analysed by sequences mapping . The aim of this study is to amplify human VKOR gene using PCR for use in cloning and expression. In this study, specific primers flanking the complete VKOR coding region was designed. The specificity of the primers were evaluated using Oligo Explorer 1.2 Software. The PCR was then performed to detect the VKOR gene. The PCR conditions were optimized by varying the PCR temperature and time. In order to prove the target PCR products were obtained after amplification, the PCR products were separated by gel electrophoresis and visualized under UV transilluminator. The band of interest which was 492 base pairs in size were obtained after human VKOR gene amplification was performed. The amplicon can then be used for cloning and expression of YKOR enzyme. However, more studies need to be performed for more specific amplification without non-specific co-amplification.
format Thesis
qualification_level Bachelor degree
author Mustafa, Nurul Ashikin
author_facet Mustafa, Nurul Ashikin
author_sort Mustafa, Nurul Ashikin
title Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa
title_short Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa
title_full Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa
title_fullStr Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa
title_full_unstemmed Amplification of human VKOR gene using PCR for use in cloning & expression / Nurul Ashikin Mustafa
title_sort amplification of human vkor gene using pcr for use in cloning & expression / nurul ashikin mustafa
granting_institution Universiti Teknologi MARA (Kampus Puncak Alam)
granting_department Faculty of Pharmacy
publishDate 2008
url https://ir.uitm.edu.my/id/eprint/100891/1/100891.PDF
_version_ 1811769105610440704