The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari

Prior to the discovery of a thermostable Taq polymerase, anyone conducting a PCR reaction procedure was obliged to sit patiently by a series of water baths or heating blocks to add a fresh aliquot of E.coli DNA polymerase after each denaturation step. This rather tedious step was eliminated by the i...

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Main Author:	Zanura, Adzhari
Format:	Student Project
Published:	Faculty of Health Science 2007
Online Access:	http://ir.uitm.edu.my/id/eprint/599/

id	uitm-599
recordtype	eprints
spelling	uitm-5992017-08-09T09:21:49Z http://ir.uitm.edu.my/id/eprint/599/ The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari Zanura, Adzhari Prior to the discovery of a thermostable Taq polymerase, anyone conducting a PCR reaction procedure was obliged to sit patiently by a series of water baths or heating blocks to add a fresh aliquot of E.coli DNA polymerase after each denaturation step. This rather tedious step was eliminated by the introduction of the Taq polymerase. Despite its huge popularity, PCR has certain limitations and the efficiency of a PCR reaction will vary according to template, reaction buffer, dNTPs, primers, cycling temperature and numbers. In this study, Taq polymerase concentrations ranging from 0.5U to 8.0U were optimized to obtain the expected DNA target. The enzyme was diluted according to the study design; standard PCR profile was then performed with other PCR components remained unchanged throughout the study. The DNA products were then electrophoresed, stained, photographed by UV light and then analyzed. At low Taq polymerase concentration (0.5U), a faint band was seen and band intensity increased proportionate to the increase of Taq polymerase. However, at 8.0U of Taq polymerase concentration, the band intensity decreased. Therefore, the optimal Taq polymerase concentration is in the range of 2.0U to 4.0U per 50ul reaction in this particular PCR protocol. It is wiser to choose a lower Taq polymerase concentration and as Taq is an expensive reagent, this may save unnecessary additional cost. In future, other alternative heat-stable DNA polymerase like Pyrocococcus furionsus (Pfii) DNA polymerase and Thermococcus litoralis (VENT) may be worth considered as an alternative because of the proof-reading ability conferred by these new polymerases. Faculty of Health Science 2007 Student Project NonPeerReviewed Zanura, Adzhari (2007) The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari. [Student Project] (Unpublished)
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Teknologi MARA
building	UiTM Institutional Repository
collection	Online Access
description	Prior to the discovery of a thermostable Taq polymerase, anyone conducting a PCR reaction procedure was obliged to sit patiently by a series of water baths or heating blocks to add a fresh aliquot of E.coli DNA polymerase after each denaturation step. This rather tedious step was eliminated by the introduction of the Taq polymerase. Despite its huge popularity, PCR has certain limitations and the efficiency of a PCR reaction will vary according to template, reaction buffer, dNTPs, primers, cycling temperature and numbers. In this study, Taq polymerase concentrations ranging from 0.5U to 8.0U were optimized to obtain the expected DNA target. The enzyme was diluted according to the study design; standard PCR profile was then performed with other PCR components remained unchanged throughout the study. The DNA products were then electrophoresed, stained, photographed by UV light and then analyzed. At low Taq polymerase concentration (0.5U), a faint band was seen and band intensity increased proportionate to the increase of Taq polymerase. However, at 8.0U of Taq polymerase concentration, the band intensity decreased. Therefore, the optimal Taq polymerase concentration is in the range of 2.0U to 4.0U per 50ul reaction in this particular PCR protocol. It is wiser to choose a lower Taq polymerase concentration and as Taq is an expensive reagent, this may save unnecessary additional cost. In future, other alternative heat-stable DNA polymerase like Pyrocococcus furionsus (Pfii) DNA polymerase and Thermococcus litoralis (VENT) may be worth considered as an alternative because of the proof-reading ability conferred by these new polymerases.
format	Student Project
author	Zanura, Adzhari
spellingShingle	Zanura, Adzhari The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari
author_facet	Zanura, Adzhari
author_sort	Zanura, Adzhari
title	The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari
title_short	The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari
title_full	The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari
title_fullStr	The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari
title_full_unstemmed	The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari
title_sort	effect of variable taq polymerase concentrations in polymerase chain reaction (pcr) / zanura adzhari
publisher	Faculty of Health Science
publishDate	2007
url	http://ir.uitm.edu.my/id/eprint/599/
first_indexed	2023-09-18T22:44:43Z
last_indexed	2023-09-18T22:44:43Z
_version_	1777417148254126080

The effect of variable taq Polymerase concentrations in Polymerase chain reaction (PCR) / Zanura Adzhari

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