Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis

Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis

A system for the study of sequence polymorphisms in animal mitochondrial DNA was developed. This system consists of two candidate markers which are mitochondrial cytochrome b and D-loop. Six novel primers were constructed for their amplifications by polymerase chain reaction (PCR). The primers K...

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Main Author: Lau, Chin Hoon
Format: Thesis
Language:English
English
Published: 1994
Subjects:
Domestic animals - Genetics
Animal breeding
DNA - Analysis
Online Access:http://psasir.upm.edu.my/id/eprint/8591/1/FSAS_1994_10_A.pdf
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spelling my-upm-ir.85912011-08-19T04:15:19Z Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis 1994 Lau, Chin Hoon A system for the study of sequence polymorphisms in animal mitochondrial DNA was developed. This system consists of two candidate markers which are mitochondrial cytochrome b and D-loop. Six novel primers were constructed for their amplifications by polymerase chain reaction (PCR). The primers KY1 and KY2 amplified a 359-bp port ion of the cytochrome b gene from chicken, water buffalo, horse, goat and sheep. Primers UPM257 and UPM258 were used to synthesize a 1.4 Kb-fragment containing the complete chicken mi tochondrial D-loop. Primers LCH4-DO and LCH2-UP paired with primers UPM257 and UPM258 respectively to amplify a 355-bp 5' region and 395-bp 3' region of the D-loop. These primers were specific in their amplifications. The mitochondrial DNA could be amplified from heterogeneous or degraded DNA preparations, as well as directly from blood incubated in proteinase K thus bypassing lengthy DNA isolation steps.Only 5ul of blood was required for a l00-ul PCR. Fort he purification of PCR products, ethanol selective precipitation was found to be both efficient and economical. Despite its lower recovery compared to the Chroma Spin column, sufficient quantity of DNA could be obtained for subsequent sequencing. Domestic animals - Genetics Animal breeding DNA - Analysis 1994 Thesis http://psasir.upm.edu.my/id/eprint/8591/ http://psasir.upm.edu.my/id/eprint/8591/1/FSAS_1994_10_A.pdf application/pdf en public masters Universiti Pertanian Malaysia Domestic animals - Genetics Animal breeding DNA - Analysis Faculty of Environmental Studies English
institution Universiti Putra Malaysia
collection PSAS Institutional Repository
language English
English
topic Domestic animals - Genetics
Animal breeding
DNA - Analysis
spellingShingle Domestic animals - Genetics
Animal breeding
DNA - Analysis
Lau, Chin Hoon
Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis
description A system for the study of sequence polymorphisms in animal mitochondrial DNA was developed. This system consists of two candidate markers which are mitochondrial cytochrome b and D-loop. Six novel primers were constructed for their amplifications by polymerase chain reaction (PCR). The primers KY1 and KY2 amplified a 359-bp port ion of the cytochrome b gene from chicken, water buffalo, horse, goat and sheep. Primers UPM257 and UPM258 were used to synthesize a 1.4 Kb-fragment containing the complete chicken mi tochondrial D-loop. Primers LCH4-DO and LCH2-UP paired with primers UPM257 and UPM258 respectively to amplify a 355-bp 5' region and 395-bp 3' region of the D-loop. These primers were specific in their amplifications. The mitochondrial DNA could be amplified from heterogeneous or degraded DNA preparations, as well as directly from blood incubated in proteinase K thus bypassing lengthy DNA isolation steps.Only 5ul of blood was required for a l00-ul PCR. Fort he purification of PCR products, ethanol selective precipitation was found to be both efficient and economical. Despite its lower recovery compared to the Chroma Spin column, sufficient quantity of DNA could be obtained for subsequent sequencing.
format Thesis
qualification_level Master's degree
author Lau, Chin Hoon
author_facet Lau, Chin Hoon
author_sort Lau, Chin Hoon
title Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis
title_short Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis
title_full Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis
title_fullStr Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis
title_full_unstemmed Development of A PCR - Cycle Sequencing System for Animal Mitochondrial DNA Analysis
title_sort development of a pcr - cycle sequencing system for animal mitochondrial dna analysis
granting_institution Universiti Pertanian Malaysia
granting_department Faculty of Environmental Studies
publishDate 1994
url http://psasir.upm.edu.my/id/eprint/8591/1/FSAS_1994_10_A.pdf
_version_ 1747810833795121152

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