Wong Ka Lok. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 121-129). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / Table A --- p.v / Table B --- p.vi / Table of Contents --- p.vii / Abbreviations --- p.xi / Chapter Chapter 1 --- Molecular authentication of endangered crocodiles and snakes / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Traditional method of snake and crocodile identification / Chapter 1.2.1 --- Morphology --- p.7 / Chapter 1.2.2 --- Chemical Analysis --- p.9 / Chapter 1.3 --- Molecular Technology in Authentication / Chapter 1.3.1 --- Polymerase Chain Reactions (PCRs) --- p.11 / Chapter 1.3.2 --- Random-primed amplification reaction --- p.12 / Chapter 1.3.3 --- Sequence Characterized Amplified Region (SCAR) --- p.13 / Chapter 1.3.4 --- PCR-RFLP --- p.13 / Chapter 1.3.5 --- DNA sequencing --- p.14 / Chapter 1.4 --- Objectives and strategies of the study --- p.15 / Chapter Chapter 2 --- Materials and General Methods / Chapter 2.1 --- Reagents and Buffers / Chapter 2.1.1 --- Buffers for Total DNA Extraction --- p.17 / Chapter 2.1.2 --- Reagents for Agarose Gel Electrophoresis --- p.17 / Chapter 2.1.3 --- Reagents for Plasmid DNA Preparation --- p.18 / Chapter 2.1.4 --- Medium for Bacterial Culture --- p.18 / Chapter 2.1.5 --- Reagents for Preparation of Competent Cells --- p.19 / Chapter 2.2 --- DNA Isolation / Chapter 2.2.1 --- Extraction of DNA from meats --- p.20 / Chapter 2.2.2 --- Extraction of DNA from blood --- p.20 / Chapter 2.3 --- Phenol/Chloroform Extraction --- p.21 / Chapter 2.4 --- Ethanol Precipitation --- p.22 / Chapter 2.5 --- DNA Concentration/Purity Estimation --- p.22 / Chapter 2.6 --- Mitochondrial DNA amplification --- p.23 / Chapter 2.7 --- Random-Primed Polymerase Chain Reactions --- p.24 / Chapter 2.8 --- SCAR for Snake samples --- p.24 / Chapter 2.9 --- SCAR for Crocodile samples --- p.25 / Chapter 2.10 --- Restriction fragment length polymorphism analysis --- p.25 / Chapter 2.11 --- Agarose Gel Electrophoresis of DNA --- p.26 / Chapter 2.12 --- Purification of PCR product --- p.26 / Chapter 2.13 --- Preparation of Escherichia coli Competent Cells --- p.27 / Chapter 2.14 --- Ligation and transformation of E. coli --- p.27 / Chapter 2.15 --- Plasmid preparation --- p.28 / Chapter 2.16 --- Screening of Plasmid DNA by Restriction Digestion --- p.29 / Chapter Chapter 3 --- DNA sequencing of snakes & construction of snake database / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Materials and methods / Chapter 3.2.1 --- Snake samples --- p.32 / Chapter 3.2.2 --- "DNA Extraction, mitochondrial gene amplification and DNA sequencing" --- p.33 / Chapter 3.2.3 --- Construction of database --- p.33 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Cytochrome b gene amplification and sequencing --- p.34 / Chapter 3.3.2 --- Gene amplification and sequencing of 16S rRNA --- p.42 / Chapter 3.3.3 --- Cytochrome b sequence database --- p.50 / Chapter 3.3.4 --- 16S rRNA sequence database --- p.53 / Chapter 3.4 --- Discussion / Chapter 3.4.1 --- Cytochrome b and 16S rRNA genes of snake species --- p.55 / Chapter 3.4.2 --- Cytochrome b and 16S rRNA databases --- p.55 / Chapter Chapter 4 --- Application of PCR-RFLP and SCAR in snake species identification / Chapter 4.1 --- Introduction --- p.57 / Chapter 4.2 --- Material and Methods / Chapter 4.2.1 --- DNA extraction and PCR-RFLP --- p.58 / Chapter 4.2.2 --- RAPD and SCAR --- p.58 / Chapter 4.3 --- Results / Chapter 4.3.1 --- PCR-RFLP of cytochrome b genes of snakes --- p.59 / Chapter 4.3.2 --- PCR-RFLP of 16S rDNA --- p.61 / Chapter 4.3.3 --- RAPD & SCAR analysis --- p.67 / Chapter 4.4 --- Discussion --- p.72 / Chapter Chapter 5 --- "Application of DNA sequencing, PCR-RFLP and SCAR to identify crocodile species" / Chapter 5.1 --- Introduction --- p.74 / Chapter 5.2 --- Materials and methods / Chapter 5.2.1 --- "Crocodile, human and four animal samples" --- p.75 / Chapter 5.2.2 --- "DNA Extraction, mitochondrial gene amplification and DNA sequencing" --- p.75 / Chapter 5.2.3 --- PCR-RFLP and SCAR --- p.76 / Chapter 5.3 --- Results / Chapter 5.3.1 --- Isolation of crocodiles DNA --- p.77 / Chapter 5.3.2 --- Isolation of DNA from Human and four animal species --- p.78 / Chapter 5.3.3 --- Cytochrome b gene amplification and sequencing --- p.78 / Chapter 5.3.4 --- 16S rRNA gene amplification and sequencing --- p.84 / Chapter 5.3.5 --- PCR-RFLP of cytochrome b --- p.89 / Chapter 5.3.6 --- PCR-RFLP of 16S rRNA --- p.91 / Chapter 5.3.7 --- SCAR primers for four crocodile species --- p.93 / Chapter 5.4 --- Discussion --- p.97 / Chapter Chapter 6 --- A case report - authentication of animal samples using DNA sequencing / Chapter 6.1 --- Introduction --- p.99 / Chapter 6.2 --- Material and methods / Chapter 6.2.1 --- Materials --- p.101 / Chapter 6.2.2 --- DNA Extraction and sequencing --- p.101 / Chapter 6.3 --- Result and discussion / Chapter 6.3.1 --- Cytochrome b gene sequencing --- p.102 / Chapter 6.3.2 --- Sequence homology among samples and meats obtained from the market --- p.111 / Chapter 6.3.3 --- Identity of samples B & D --- p.113 / Chapter Chapter 7 --- General Discussion / Chapter 7.1 --- Advantages and weakness of DNA technology --- p.116 / Chapter 7.2 --- Choosing appropriate molecular markers --- p.118 / Chapter 7.3 --- Further suggested work --- p.119 / Chapter 7.4 --- Conclusion --- p.119 / References --- p.121 / Appendix --- p.130