Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]

The polymerase chain reaction (PCR) was an essential fundamental procedure widely used in molecular biology to amplify specific deoxyribonucleic acid (DNA) sequences. To investigate the efficiency and specificity of PCR using DNA extracted from human blood cells as sample, several concentrations of...

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Main Authors: Mazlan, Alif Haikal, Muhamad Najib, Muhamad Hafizal Aqhmal, Hazizul Hassan, Mizaton, Mohd Hatta, Fazleen Haslinda, Mohd Yusoff, Rosmadi
Format: Article
Language:English
Published: Faculty of Pharmacy 2024
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Online Access:https://ir.uitm.edu.my/id/eprint/94561/1/94561.pdf
https://ir.uitm.edu.my/id/eprint/94561/
http://ijpncs.uitm.edu.my/index.php/en/ijpncs-journal
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Institution: Universiti Teknologi Mara
Language: English
id my.uitm.ir.94561
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spelling my.uitm.ir.945612024-05-30T14:54:36Z https://ir.uitm.edu.my/id/eprint/94561/ Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.] ijpnacs Mazlan, Alif Haikal Muhamad Najib, Muhamad Hafizal Aqhmal Hazizul Hassan, Mizaton Mohd Hatta, Fazleen Haslinda Mohd Yusoff, Rosmadi Medical education. Medical schools. Research The polymerase chain reaction (PCR) was an essential fundamental procedure widely used in molecular biology to amplify specific deoxyribonucleic acid (DNA) sequences. To investigate the efficiency and specificity of PCR using DNA extracted from human blood cells as sample, several concentrations of DNA templates were used to obtain accurate and reproducible results during this optimisation process. The principal objective was to identify which concentration was the best for amplifying the DNA target sequence and study the effects of the scale of concentrations to standard PCR. For this study, a series of PCR were conducted using various concentrations of DNA template ranging from 1 ng/μL to 100 ng/μL per reaction under optimized conditions of primer concentrations, annealing temperature and extension time. The results demonstrated that DNA template concentration significantly influenced the efficiency and specificity of PCR as the intensity of the target gene amplification band was increased on the agarose gel electrophoresis under ultraviolet light indicating the PCR product yield. Formation of primer-dimers became more prominent as the DNA template concentration reduced resulting in non-specific amplification. The concentrations ranging from 10 ng/µL to 70 ng/µL were the most optimal reactions profiting a remarkable feat of target gene amplification while concentrations less than 1 ng/µL produced none and non-specific outcomes retaining little amount of target gene amplification. In conclusion, our study highlights the pivotal role of DNA template concentration optimises PCR to increase reliability and reproducibility, extending our understanding of genetic analysis, diagnostic, and forensic sciences. Faculty of Pharmacy 2024 Article PeerReviewed text en https://ir.uitm.edu.my/id/eprint/94561/1/94561.pdf Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]. (2024) International Journal of Pharmaceutical, Nutraceutical and Cosmetic Science (IJPNaCS) <https://ir.uitm.edu.my/view/publication/International_Journal_of_Pharmaceutical,_Nutraceutical_and_Cosmetic_Science_=28IJPNaCS=29/>, 7 (1). pp. 1-11. ISSN 2682-8146 http://ijpncs.uitm.edu.my/index.php/en/ijpncs-journal
institution Universiti Teknologi Mara
building Tun Abdul Razak Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Mara
content_source UiTM Institutional Repository
url_provider http://ir.uitm.edu.my/
language English
topic Medical education. Medical schools. Research
spellingShingle Medical education. Medical schools. Research
Mazlan, Alif Haikal
Muhamad Najib, Muhamad Hafizal Aqhmal
Hazizul Hassan, Mizaton
Mohd Hatta, Fazleen Haslinda
Mohd Yusoff, Rosmadi
Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]
description The polymerase chain reaction (PCR) was an essential fundamental procedure widely used in molecular biology to amplify specific deoxyribonucleic acid (DNA) sequences. To investigate the efficiency and specificity of PCR using DNA extracted from human blood cells as sample, several concentrations of DNA templates were used to obtain accurate and reproducible results during this optimisation process. The principal objective was to identify which concentration was the best for amplifying the DNA target sequence and study the effects of the scale of concentrations to standard PCR. For this study, a series of PCR were conducted using various concentrations of DNA template ranging from 1 ng/μL to 100 ng/μL per reaction under optimized conditions of primer concentrations, annealing temperature and extension time. The results demonstrated that DNA template concentration significantly influenced the efficiency and specificity of PCR as the intensity of the target gene amplification band was increased on the agarose gel electrophoresis under ultraviolet light indicating the PCR product yield. Formation of primer-dimers became more prominent as the DNA template concentration reduced resulting in non-specific amplification. The concentrations ranging from 10 ng/µL to 70 ng/µL were the most optimal reactions profiting a remarkable feat of target gene amplification while concentrations less than 1 ng/µL produced none and non-specific outcomes retaining little amount of target gene amplification. In conclusion, our study highlights the pivotal role of DNA template concentration optimises PCR to increase reliability and reproducibility, extending our understanding of genetic analysis, diagnostic, and forensic sciences.
format Article
author Mazlan, Alif Haikal
Muhamad Najib, Muhamad Hafizal Aqhmal
Hazizul Hassan, Mizaton
Mohd Hatta, Fazleen Haslinda
Mohd Yusoff, Rosmadi
author_facet Mazlan, Alif Haikal
Muhamad Najib, Muhamad Hafizal Aqhmal
Hazizul Hassan, Mizaton
Mohd Hatta, Fazleen Haslinda
Mohd Yusoff, Rosmadi
author_sort Mazlan, Alif Haikal
title Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]
title_short Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]
title_full Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]
title_fullStr Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]
title_full_unstemmed Effect of DNA template concentration on standard polymerase chain reaction / Alif Haikal Mazlan ... [et al.]
title_sort effect of dna template concentration on standard polymerase chain reaction / alif haikal mazlan ... [et al.]
publisher Faculty of Pharmacy
publishDate 2024
url https://ir.uitm.edu.my/id/eprint/94561/1/94561.pdf
https://ir.uitm.edu.my/id/eprint/94561/
http://ijpncs.uitm.edu.my/index.php/en/ijpncs-journal
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