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課程名稱 |
生物技術核心實驗
Biotechnology Core Techniques |
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開課學期 |
106-1 |
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授課對象 |
生命科學系 |
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授課教師 |
張英峯 |
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課號 |
Prog5074 |
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課程識別碼 |
P05 U2020 |
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班次 |
02 |
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學分 |
4.0 |
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全/半年 |
半年 |
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必/選修 |
必修 |
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上課時間 |
星期四2,3,4,5,6,7,8,9,10(9:10~18:20) |
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上課地點 |
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備註 |
初選不開放。生科系優先登記。上課教室：生科館3C教室。外系需繳學分費。與鄭貽生、朱家瑩合授
總人數上限：40人 |
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Ceiba 課程網頁 |
http://ceiba.ntu.edu.tw/1061Prog5074_02 |
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課程簡介影片 |
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核心能力關聯 |
本課程尚未建立核心能力關連 |
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課程大綱
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為確保您我的權利,請尊重智慧財產權及不得非法影印
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課程概述 |
本課程為生物技術學程之重要基礎實驗科目，整個流程以分子生物中心法則(Central Dogma)為骨幹，學習由GUS酵素基因之DNA開始，經過剪接與基因轉殖，從宿主細胞中表現並檢定其mRNA，然後收集純化並檢定目標酵素。
1. DNA manipulation
(1). Basic Techniques
a. Pipetting
b. Bacterial culture techniques
(2). DNA preparation
a. Miniprep of plasmid DNA
b. Restriction analysis
c. Large scale digestion of plasmid
(3). DNA ligation
a. Isolation of DNA fragment
b. Quantification analysis of DNA
c. DNA ligation
(4). Transformation
a. Transformation by CaCl2
(5). Protein expression system
a. Colony hybridization
b. Histochemical detection
(6). Detection of specific nucleic acid sequences by hybridization
a. mini-prep of plasmid DNA
b. Restriction analysis
c. Southern boltting
2. RNA hybridization
(1). RNA Preparation
a.Total RNA isolation
b.In vitro transcription
(2). DIG-labeled probe preparation
a.Random priming
b.PCR reaction
(3). Northern blot hybridization
a. RNA analysis
b. Northern blotting
c. Northern blot hybridization
d. Detection of probe by enzyme immunoassay
e. Analysis of the blot
3.Protein analysis
(1). Protein purification
a. Protein extraction
b. Gel filtration
(2). Ion exchange chromatography
a. Ion exchange chromatography
b. Activity assay
(3). Affinity chromatography
a. Affinity chromatography
b. Activity assay
(4). Electrophoresis analysis
a. SDS-PAGE
b. Protein transfer
c. Immunostaining |
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課程目標 |
本課程由基本的自動吸管開始，教導基礎的核酸及蛋白質操作技術，對出入生物技術領域的初學者有很大的幫助。
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課程要求 |
本課程共分成DNA、RNA、蛋白質三大部份，分別由三位教師分別授課。
成績計算方式為；依照各部份小考、段考以及報告成績計算出各段成績，再以三段成績及平時操作表現計算出總成績。 |
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預期每週課後學習時數 |
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Office Hours |
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指定閱讀 |
待補 |
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參考書目 |
Adams RLP, Knowler JT & Leader DP (1992) The Biochemistry of the Nucleic Acids. 11th ed. Chapman & Hall Ltd. 675pp.
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA & Struhl K (2004) Current Protocols in Molecular Biology. John Wiley & Sons, Inc. Chichester, UK.
Blackburn GM & Gait MJ (1996) Nucleic Acids in Chemistry and Biology. 2nd ed. Oxford University Press Inc., New York. 528pp.
Brown TA (2010) Gene Cloning and DNA Analysis. 6th ed. Blackwell Science Ltd, London. 363pp.
Clark DP (2005) Molecular Biology - Understanding the Genetic Revolution. Elsevier Academic Press, USA. 784pp.
Glover DM (1984) Gene Cloning. Chapman and Hall, London, New York. 222pp.
Micklos DA, Freyer GA & Crotty DA (2003) DNA Science – A First Course. Cold Spring Harbor Laboratory Press. 575pp.
Miesfeld RL (1999) Applied Molecular Genetics. Wiley-Liss, Inc. 293pp.
Primrose SB, Twyman RM & Old RW (2001) Principles of Gene Manipulation. Blackwell Science Ltd, London. 390pp.
Sambrook J & Russell DW (2001) Molecular Cloning – A Laboratory Manual. 3rd ed. Vols 1, 2 & 3. Cold Spring Harbor Laboratory Press, New York.
Spangler BD (2002) Methods in Molecular Biology and Protein Chemistry. John Wiley & Sons, Ltd., Chichester, UK. 209pp.
Summers DK (1996) The Biology of Plasmids. Blackwell Science Ltd. 157pp.
Tagu D & Moussard C (2006) Techniques for Molecular Biology. Science Publishers, Enfield, NH, USA. 227pp.
Tropp BE (2008) Molecular Biology – Genes to Proteins. Jones and Bartlett Publishers, USA. 1000pp.
Walker MR & Rapley R (1997) Route Maps in Gene Technology. Blackwell Science Ltd. 334pp.
Watson JD, Caudy AA, Myers RM & Witkowski JA (2007) Recombinant DNA. 3rd ed. Cold Spring Harbor Laboratory Press, New York. 474pp. |
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評量方式
(僅供參考) |
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週次 |
日期 |
單元主題 |
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第1週 |
9/14 |
【DNA-0】Registration |
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第2週 |
9/21 |
【DNA-1】Introduction and cloning tools |
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第3週 |
9/28 |
【DNA-2】Cloning strategies |
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第4週 |
10/05 |
【DNA3】 Cloning strategies (II): Restriction analysis and ligation |
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第5週 |
10/12 |
【DNA-4】Transformation and identification of transformants |
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第6週 |
10/19 |
【DNA-5】Protein expression systems |
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第7週 |
10/26 |
【DNA-6】Detection of specific nucleic acid sequences by hybridization |
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第8週 |
11/02 |
DNA Examination, DNA report |
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第9週 |
11/09 |
【RNA-1】RNA in general and RNA preparation |
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第10週 |
11/16 |
【RNA-2】Denaturing gel electrophoresis and probe preparation |
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第11週 |
11/23 |
【RNA-3】Northern hybridization analysis |
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第12週 |
11/30 |
【RNA-4】Analysis of RNA by primer extension, S1 analysis, RNase protection, nuclear run-off, and RT-PCR |
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第13週 |
12/07 |
【Protein-1】Protein in general and protein extraction |
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第14週 |
12/14 |
【Protein-2】Ion exchange chromatography and Affinity chromatography |
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第15週 |
12/21 |
【Protein-3】Protein assay Polyacrylamide electrophoresis Protein transfer |
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第16週 |
12/28 |
【Protein-4】Immunological tools |
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第17週 |
1/04 |
Protein Examination, Experiment Examination, Protein report |
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