Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor

Background: The DNA polymerase from Pyrococcus furiosus (Pfu) has gained considerable attention in the field of DNA amplification as this enzyme have elevated temperature optima and have thermal stabilities that roughly correspond to the thermal extremes of the environment from which they were isola...

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Main Author:	Hasnor, Sayeeda Zainab
Format:	Thesis
Language:	English
Published:	2009
Subjects:	Pharmaceutical industry Pharmaceutical chemistry
Online Access:	https://ir.uitm.edu.my/id/eprint/105400/1/105400.PDF
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id	my-uitm-ir.105400
record_format	uketd_dc
spelling	my-uitm-ir.1054002024-12-04T16:03:05Z Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor 2009 Hasnor, Sayeeda Zainab Pharmaceutical industry Pharmaceutical chemistry Background: The DNA polymerase from Pyrococcus furiosus (Pfu) has gained considerable attention in the field of DNA amplification as this enzyme have elevated temperature optima and have thermal stabilities that roughly correspond to the thermal extremes of the environment from which they were isolated. Due to thermostability of this enzyme, the structure and function relationships and the potential industrial applications of many thermostable enzymes such as DNA polymerases are of considerable interest to researchers. Aim: The purpose of this study is to clone the Recombinant DNA polymerase from Pyrococcus furiosus in Escherichia coli. Method: The PCR was conducted based on standard PCR protocol and was optimized under different annealing temperatures using EppendorfMastercycler Gradient to determine the optimal annealing temperature for this method. And also for further optimization, new primers batch was developed to eliminate contaminate primers. Results: The PCR based method was failed to give the result. It shows that there was no DNA amplification but only DNA ladder band can be seen. Conclusion: This study failed to amplify the Pfu DNA polymerase gene. More time needed to optimize the PCR condition. 2009 Thesis https://ir.uitm.edu.my/id/eprint/105400/ https://ir.uitm.edu.my/id/eprint/105400/1/105400.PDF text en public degree Universiti Teknologi MARA (Kampus Puncak Alam) Faculty of Pharmacy Lee, Wee Leng
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Lee, Wee Leng
topic	Pharmaceutical industry Pharmaceutical chemistry
spellingShingle	Pharmaceutical industry Pharmaceutical chemistry Hasnor, Sayeeda Zainab Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor
description	Background: The DNA polymerase from Pyrococcus furiosus (Pfu) has gained considerable attention in the field of DNA amplification as this enzyme have elevated temperature optima and have thermal stabilities that roughly correspond to the thermal extremes of the environment from which they were isolated. Due to thermostability of this enzyme, the structure and function relationships and the potential industrial applications of many thermostable enzymes such as DNA polymerases are of considerable interest to researchers. Aim: The purpose of this study is to clone the Recombinant DNA polymerase from Pyrococcus furiosus in Escherichia coli. Method: The PCR was conducted based on standard PCR protocol and was optimized under different annealing temperatures using EppendorfMastercycler Gradient to determine the optimal annealing temperature for this method. And also for further optimization, new primers batch was developed to eliminate contaminate primers. Results: The PCR based method was failed to give the result. It shows that there was no DNA amplification but only DNA ladder band can be seen. Conclusion: This study failed to amplify the Pfu DNA polymerase gene. More time needed to optimize the PCR condition.
format	Thesis
qualification_level	Bachelor degree
author	Hasnor, Sayeeda Zainab
author_facet	Hasnor, Sayeeda Zainab
author_sort	Hasnor, Sayeeda Zainab
title	Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor
title_short	Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor
title_full	Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor
title_fullStr	Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor
title_full_unstemmed	Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor
title_sort	cloning of thermostable recombinant dna polymerase in escherichia coli / sayeeda zainab hasnor
granting_institution	Universiti Teknologi MARA (Kampus Puncak Alam)
granting_department	Faculty of Pharmacy
publishDate	2009
url	https://ir.uitm.edu.my/id/eprint/105400/1/105400.PDF
_version_	1818588147063193600

Cloning of thermostable recombinant dna polymerase in escherichia coli / Sayeeda Zainab Hasnor

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