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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 盧彥文 | |
| dc.contributor.author | Kan-Chien Li | en |
| dc.contributor.author | 李侃謙 | zh_TW |
| dc.date.accessioned | 2021-06-07T23:58:11Z | - |
| dc.date.copyright | 2013-09-25 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-17 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17142 | - |
| dc.description.abstract | 單核苷酸多型性 (Single Nucleotide Polymorphism, SNP)為去氧核醣核酸(DNA)的序列中發生一個鹼基對的變異。這類的變異若發生在編碼區則有可能引起蛋白質氨基酸編碼改變進而影響原本正常功能的表現。檢測這類的基因變異，去氧核醣核酸熔解曲線(DNA熔解曲線)的檢測方式是一個非常經濟且有效的手段。近年來，許多研究將微流體平台大量應用於熔解曲線的分析，主因於其技術不但可以大幅減少使用試劑，並且能微小化檢測平台，成為重點照護的檢測裝置。目前以微流體檢測DNA熔解曲線研究中，為了避免檢體在前處理及檢測時交叉污染，只能以單次檢測單項、固定同一檢體於個別區域等方式進行低通量的檢測。為了改善這些限制，本研究以毛細血管擴張性運動失調突變基因(ataxia-telangiectasia mutated, ATM) 為例，將個別檢體固定於不同群體的微珠上，並且循序注入內建微溫度場的微流道中進行基因檢測，成功由藍瑞斯母豬取得之基因體DNA完成位點ATM-A的基因分型。 | zh_TW |
| dc.description.abstract | Deoxyribonucleic acid (DNA) melting analysis is a powerful tool in genotyping and mutation scanning. The recent development in microfluidics further promotes melting analysis within a miniaturized device, which not only can reduce the reagent cost but also provide the possibility to perform point-of-care molecular diagnosis. The DNA melting analysis in microfluidic devices usually resorts to solid or liquid phase on sample preparations, in which either DNA immobilization on channel surfaces is required or only single analysis is allowed. To address these limitations, a bead-based melting analysis in continuous flow configuration is proposed. Our device is tested on detecting single nucleotide polymorphisms (SNPs) - one of the most important DNA sequence variations for human, animals and other species. The samples with ataxia telangiectasia-mutated (ATM) genes from Landrace sows, which play important roles in total number of piglets born, number born alive and average birth weight due to its differential expression between the morula and blastocyst stages, were chosen as our SNP genotyping candidates. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-07T23:58:11Z (GMT). No. of bitstreams: 1 ntu-102-R00631034-1.pdf: 3360746 bytes, checksum: 479d1c2bb1b73176fa14274271e8cdd5 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 誌謝 i
中文摘要 ii Abstract iii List of Figures vii List of Tables xii Chapter 1 Introduction 1 1.1 Single Nucleotide Polymorphism and Ataxia-Telangiectasia Mutate 1 1.2 Microfluidic Technology 2 1.3 Overall Structure of Thesis 3 Chapter 2 Literature Review 4 2.1 Single Nucleotide Polymorphism detection Strategies 5 2.2 Melting Curve Analysis for Genotyping 9 2.3 Bead-Based Microfluidics Device 12 2.4 Integrated Temperature Control System with Microfluidic Device 14 Chapter 3 Materials and Methods 16 3.1 ATM Gene Preparation 17 3.1.1 Symmetric Polymerase Chain Reaction 18 3.1.2 Asymmetric Polymerase Chain Reaction 20 3.2 Device Fabrication 22 3.2.1 Fabrication for Microchannel 22 3.2.2 Fabrication for ITO Heater and Thermometers 25 3.3 Chip Design 27 3.3.1 Microchannel Design 27 3.3.2 Heater and Thermometer Design 29 3.3.3 Temperature measurement 30 3.3.4 Temperature Control Method 31 3.4 Bead-based Melting Strategies 32 3.4.1 Melting Protocol 34 3.5 Data Acquisition on Fluorescent Images 38 3.5.1 Melting Curve Analysis 39 3.6 Genotyping Validation 40 Chapter 4 Results and Discussion 41 4.1 Temperature Calibration 42 4.2 Temperature Control System 45 4.3 Microbead Manipulation Experiment 48 4.4 Melting Curve Analysis 51 4.4.1 Melting curve analysis by NTU SNP detection system 51 4.4.2 Melting Curve Analysis by Rotor-Gene Q System 53 4.4.3 Comparisons of the bead-based SNP Detection System and Rotor-Gene Q System 55 4.4.4 Relationship between Heating Rate and Melting Temperature 59 Chapter 5 Conclusions 63 5.1 Conclusions 63 5.2 Future Prospects 64 Appendix I 66 Appendix II 69 Appendix III 71 Appendix IV 74 Reference 76 | |
| dc.language.iso | en | |
| dc.title | 利用微珠與微流道溫度梯度場分析熔解曲線進行單核苷酸多態性檢測 | zh_TW |
| dc.title | Bead-based Melting Analysis in Temperature-gradient Microchannel for Single Nucleotide Polymorphism Detection | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 丁詩同,王倫,林恩仲,范士岡 | |
| dc.subject.keyword | 微珠,微流道,單核?酸多型性,氧核醣核酸熔解曲線分析,毛細血管擴張性運動失調突變基因, | zh_TW |
| dc.subject.keyword | microfluidics,single nucleotide polymorphisms,ataxia telangiectasia-mutated,bead-based melting analysis, | en |
| dc.relation.page | 68 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2013-08-17 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物產業機電工程學研究所 | zh_TW |
| 顯示於系所單位： | 生物機電工程學系 | |
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