Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali

Polymerase Chain Reaction (PCR) is typically carried om in three distinct steps governed by three distinct temperatures. These three important steps are the denaturing, annealing and extension. Annealing is the second stage of a PCR run whereby the primers will bind to the target DNA or target seque...

Full description

Saved in:

Bibliographic Details
Main Author:	Ali, Mohd Khairul Shafiq
Format:	Thesis
Language:	English
Published:	2007
Subjects:	Public health Hygiene Preventive Medicine Health behavior and habits
Online Access:	https://ir.uitm.edu.my/id/eprint/39733/1/39733.pdf
Tags:	Add Tag No Tags, Be the first to tag this record!

id	my-uitm-ir.39733
record_format	uketd_dc
spelling	my-uitm-ir.397332021-07-23T16:56:45Z Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali 2007 Ali, Mohd Khairul Shafiq Public health. Hygiene. Preventive Medicine Health behavior and habits Polymerase Chain Reaction (PCR) is typically carried om in three distinct steps governed by three distinct temperatures. These three important steps are the denaturing, annealing and extension. Annealing is the second stage of a PCR run whereby the primers will bind to the target DNA or target sequence for amplification. The goal is not to amplify in vitro the entire DNA but only specific region of interest only. In this study, the effects of variable or different annealing temperature on PCR performance were investigated. Six different annealing temperature setting were chosen, that is 46° C, 49°C, ,52°C, 55°C, 58°C, and 61°C. At the end of the PCR run the amplicons were then electrophoresed, stained with ethidium bromide dammed in water and finally photographed by a gel documentation system with UV light. At 46°C to 58°C DNA bands of varying intensity were produced. At 61°C, no band was seen. These results showed that the best annealing temperature (Tm) for this consensus primer was 52°C. The band was clear, sharp and with the highest intensity at the expected 1400 bp molecular weight. This therefore concludes that the Tm of a particular primer sets (forward and reverse) can be experimentally determined to obtain the optimum result. 2007 Thesis https://ir.uitm.edu.my/id/eprint/39733/ https://ir.uitm.edu.my/id/eprint/39733/1/39733.pdf text en public degree Universiti Teknologi MARA Faculty of Health Sciences Hassan, Zuridah (Dr.)
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Hassan, Zuridah (Dr.)
topic	Public health Hygiene Preventive Medicine Health behavior and habits
spellingShingle	Public health Hygiene Preventive Medicine Health behavior and habits Ali, Mohd Khairul Shafiq Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali
description	Polymerase Chain Reaction (PCR) is typically carried om in three distinct steps governed by three distinct temperatures. These three important steps are the denaturing, annealing and extension. Annealing is the second stage of a PCR run whereby the primers will bind to the target DNA or target sequence for amplification. The goal is not to amplify in vitro the entire DNA but only specific region of interest only. In this study, the effects of variable or different annealing temperature on PCR performance were investigated. Six different annealing temperature setting were chosen, that is 46° C, 49°C, ,52°C, 55°C, 58°C, and 61°C. At the end of the PCR run the amplicons were then electrophoresed, stained with ethidium bromide dammed in water and finally photographed by a gel documentation system with UV light. At 46°C to 58°C DNA bands of varying intensity were produced. At 61°C, no band was seen. These results showed that the best annealing temperature (Tm) for this consensus primer was 52°C. The band was clear, sharp and with the highest intensity at the expected 1400 bp molecular weight. This therefore concludes that the Tm of a particular primer sets (forward and reverse) can be experimentally determined to obtain the optimum result.
format	Thesis
qualification_level	Bachelor degree
author	Ali, Mohd Khairul Shafiq
author_facet	Ali, Mohd Khairul Shafiq
author_sort	Ali, Mohd Khairul Shafiq
title	Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali
title_short	Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali
title_full	Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali
title_fullStr	Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali
title_full_unstemmed	Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali
title_sort	effect of variable annealing temperatures in polymerase chain reaction / mohd khairul shafiq ali
granting_institution	Universiti Teknologi MARA
granting_department	Faculty of Health Sciences
publishDate	2007
url	https://ir.uitm.edu.my/id/eprint/39733/1/39733.pdf
_version_	1783734531734372352

Effect of variable annealing temperatures in Polymerase Chain Reaction / Mohd Khairul Shafiq Ali

Similar Items