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Arbitrarily primed polymerase chain reaction and electrophoretic karyotype analyses of Shiitake mushroom (Lentinula edodes).

by Lai, Shiu Hong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 129-147). / TITLE PAGE --- p.I / THESIS COMMITTEE --- p.II / ABSTRACT --- p.III / ACKNOWLEDGMENTS --- p.V / ABBREVIATIONS --- p.VI / TABLE OF CONTENTS --- p.VII / LIST OF TABLES --- p.XI / LIST OF FIGURES --- p.XII / Chapter Chapter 1. --- Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) Analysis of Lent inula edodes / Chapter 1. --- Introduction / Chapter 1.1 --- General Introduction --- p.1 / Chapter 1.2 --- Purpose of Study --- p.4 / Chapter 2. --- Literature Review / Chapter 2.1 --- Biology of Lentinula edodes / Chapter 2.1.1 --- "Overview," --- p.6 / Chapter 2.1.2 --- Life Cycle of Lentinula edodes --- p.6 / Chapter 2.1.3 --- Dedikaryotization (Monokaryotization) --- p.12 / Chapter 2.2 --- Genome Analysis of the Mushroom --- p.13 / Chapter 2.3 --- Genetic Markers of Lentinula edodes / Chapter 2.3.1 --- Overview --- p.15 / Chapter 2.3.2 --- Auxotrophic Markers --- p.16 / Chapter 2.3.3 --- Biochemical Markers --- p.17 / Chapter 2.3.4 --- Molecular Markers / Chapter 2.3.4.1 --- RFLPs --- p.19 / Chapter 2.3.4.2 --- PCR-Based Markers --- p.20 / Chapter 2.4 --- Polymerase Chain Reaction (PCR) / Chapter 2.4.1 --- The principle of PCR --- p.22 / Chapter 2.4.2 --- Applications of PCR on Mushroom Studies --- p.26 / Chapter 2.5 --- Arbitrarily Primed Polymerase Chain Reaction / Chapter 2.5.1 --- Principle of AP-PCR --- p.27 / Chapter 2.5.2 --- Applications of AP-PCR on Mushroom Studies --- p.29 / Chapter 2.6 --- Genetic Linkage Analysis / Chapter 2.6.1 --- Overview --- p.31 / Chapter 2.6.2 --- The LOD Score Method --- p.34 / Chapter 3. --- Materials and Methods / Chapter 3.1 --- Mushroom Strains and Culture Media --- p.36 / Chapter 3.2 --- Culture Method --- p.36 / Chapter 3.3 --- Solutions --- p.36 / Chapter 3.4 --- Primers --- p.38 / Chapter 3.5 --- Isolation of DNA from Lentinula edodes / Chapter 3.5.1 --- Mini-Preparation of Fungal DNA from L. edodes for PCR amplification --- p.42 / Chapter 3.5.2 --- Cesium Chloride Method: Mini-Preparation of Fungal DNA for PCR amplification --- p.43 / Chapter 3.6 --- Quantitative Measurements of DNA --- p.44 / Chapter 3.7 --- Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) for the Amplification of Genomic DNA of L. edodes --- p.45 / Chapter 3.8 --- Analysis of DNA Samples with Agarose Gel Electrophoresis --- p.46 / Chapter 3.9 --- Analysis of DNA Samples with Polyacrylamide Gel Electrophoresis (PAGE) --- p.47 / Chapter 3.10 --- Silver Staining --- p.48 / Chapter 3.11 --- Single Stranded Conformation Polymorphism (SSCP) Analysis of Polymorphic DNA Fragments / Chapter 3.11.1 --- Elution and Amplification of DNA --- p.49 / Chapter 3.11.2 --- PCR-SSCP --- p.50 / Chapter 3.12 --- Segregation and Linkage Analysis / Chapter 3.12.1 --- Chi-Square Test --- p.51 / Chapter 3.12.2 --- The LOD Score Method --- p.52 / Chapter 4. --- Results / Chapter 4.1 --- DNA Extraction --- p.54 / Chapter 4.2 --- AP-PCR Amplified Fragments and Fragment Number --- p.58 / Chapter 4.3 --- Dedikaryotization Demonstration --- p.60 / Chapter 4.4 --- Identification of Polymorphic Genetic Markers --- p.64 / Chapter 4.4.1 --- AP-PCR Fingerprints from Single Primer --- p.66 / Chapter 4.4.2 --- AP-PCR Fingerprints Using Two Primers --- p.76 / Chapter 4.5 --- Segregation of Polymorphic Markers in Single Spore Isolates (SSIs) --- p.81 / Chapter 4.6 --- Single Stranded Conformation Polymorphism (SSCP) of Identified Polymorphic DNA Fragments --- p.86 / Chapter 4.7 --- Linkage Analysis of the Identified AP-PCR Markers --- p.89 / Chapter 5. --- Discussions / Chapter 5.1 --- DNA Extraction --- p.92 / Chapter 5.2 --- Arbitrary Primers --- p.93 / Chapter 5.3 --- Dedikaryotization Demonstration --- p.95 / Chapter 5.4 --- Identification of Polymorphic Genetic Markers --- p.96 / Chapter 5.5 --- AP-PCR Analysis of a Mushroom --- p.97 / Chapter 5.6 --- Mendelian Segregation Pattern of the Polymorphic Markers --- p.99 / Chapter 6. --- Conclusion and Further Studies --- p.101 / Chapter 2. Electrophoretic Karyotype Analysis of Lentinula edodes / Chapter 7. --- Introduction --- p.106 / Chapter 8. --- Literature Review / Chapter 8.1 --- Overview --- p.108 / Chapter 8.2 --- Protoplasts --- p.109 / Chapter 8.3 --- Pulsed Field Gel Electrophoresis (PFGE) / Chapter 8.3.1 --- Principle --- p.110 / Chapter 8.3.2 --- Applications of PFGE in Studies of Fungi --- p.112 / Chapter 9. --- Materials and Methods / Chapter 9.1 --- Strains and Culture Media --- p.114 / Chapter 9.2 --- Solutions --- p.114 / Chapter 9.3 --- Production of Lentinula edodes Protoplast --- p.115 / Chapter 9.4 --- Electrophoretic Conditions --- p.116 / Chapter 9.4.1 --- Condition for Saccharomyces cerevisiae chromosomes --- p.117 / Chapter 9.4.2 --- Condition for Candida albicans chromosomes --- p.117 / Chapter 9.4.3 --- Condition for Schizosaccharomyces pombe chromosomes --- p.118 / Chapter 10. --- Results / Chapter 10.1 --- Protoplast Production of Lentinula edodes / Chapter 10.1.1 --- Effects of Age of Mycelium on Protoplast Yield --- p.119 / Chapter 10.1.2 --- Effects of Various Osmotic Stabilizers on Protoplast Yield --- p.121 / Chapter 10.1.3 --- Effects of Two Lytic Enzymes on Protoplast Yield --- p.123 / Chapter 10.1.4 --- The Optimal Condition --- p.123 / Chapter 10.2 --- Electrophoretic Karyotype of L. edodes --- p.124 / Chapter 11. --- Discussions / Chapter 11.1 --- Protoplast Production of Lentinula edodes --- p.126 / Chapter 11.2 --- Electrophoretic Karyotype --- p.127 / REFERENCES

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_319130
Date January 1993
ContributorsLai, Shiu Hong., Chinese University of Hong Kong Graduate School. Division of Biology.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xiv, 147 leaves : ill. (chiefly mounted) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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