《Biochemistry Experiment V)》课程教学大纲 一、课程基本信息 英文名称 (Biochemistry Experiment V) 课程代码 PHAR1114 课程性质 大类基础课程 授课对象 药学全英文班 学 分 学 时 36 Faculty from Department of 主讲教师 修订日期 2021年6月1日 Biochemistry and Molecular Biology Experimental Biochemistry essentials 指定教材 生物化学与分子生物学实验教程徐岚、钱晖科学出版社、2014 二、课程目标 (一）总体目标 (以三维目标即知识与技能、过程与方法、情感态度与价值观的形式反映核心素养观念 和内容，其中核心素养不仅关注学生“当下发展”，更关注学生“未来发展”所需要的正确 价值观念、必备品格和关键能力，即把知识、技能和过程、方法提炼为能力，把情感态度、 价值观提炼为品格) The topics and experiments in this Laboratory complement the material covered in the course of Biochemistry and Molecular Biology.This course provides an introduction to basic laboratory techniques and experimental methods,as well as direct experience with biochemistry and molecular biology principles and the properties and reactions of biomolecules. Students attend lab lectures and perform experiments every week as scheduled. This course includes 9 sessions (4 hours/session).In addition to the basic training in laboratory techniques,it covers analyses of nucleotides,proteins,lipids and other metabolites and measurement of enzymatic activities.This course also includes 6 comprehensive experiments,which help students prepare for future work in a research lab. (二)课程目标
《Biochemistry Experiment V》课程教学大纲 一、课程基本信息 英文名称 （Biochemistry Experiment V） 课程代码 PHAR1114 课程性质 大类基础课程 授课对象 药学全英文班 学 分 1 学 时 36 主讲教师 Faculty from Department of Biochemistry and Molecular Biology 修订日期 2021 年 6 月 1 日 指定教材 Experimental Biochemistry Essentials 生物化学与分子生物学实验教程 徐岚、钱晖 科学出版社、2014 二、课程目标 （一）总体目标 （以三维目标即知识与技能、过程与方法、情感态度与价值观的形式反映核心素养观念 和内容，其中核心素养不仅关注学生“当下发展”，更关注学生“未来发展”所需要的正确 价值观念、必备品格和关键能力，即把知识、技能和过程、方法提炼为能力，把情感态度、 价值观提炼为品格） The topics and experiments in this Laboratory complement the material covered in the course of Biochemistry and Molecular Biology. This course provides an introduction to basic laboratory techniques and experimental methods, as well as direct experience with biochemistry and molecular biology principles and the properties and reactions of biomolecules. Students attend lab lectures and perform experiments every week as scheduled. This course includes 9 sessions (4 hours/session). In addition to the basic training in laboratory techniques, it covers analyses of nucleotides, proteins, lipids and other metabolites and measurement of enzymatic activities. This course also includes 6 comprehensive experiments, which help students prepare for future work in a research lab. （二）课程目标
(课程目标规定某一阶段的学生通过课程学习以后，在发展德、智、体、美、劳等方面 期望实现的程度，它是确定课程内容、教学目标和教学方法的基础。) 课程目标l:Help students to recapitulate the basic knowledge covered in the course of Biochemistry and Molecular biology. 1.1 Help students to recapitulate the basic knowledge learnt from the theoretical class 1.2 Help students to understand the principles of several basic biochemistry experiments. 课程目标2：Prepare students for the future work in research labs 2.1 Training student the basic biochemistry experimental skills 2.2 Help students to learn lab report writing skills (要求参照《普通高等学校本科专业类教学质量国家标准》，对应各类专业认证标准， 注意对毕业要求支撑程度强弱的描述，与“课程目标对毕业要求的支撑关系表一致)（五号 宋体) （三)课程目标与毕业要求、课程内容的对应关系 表1：课程目标与课程内容、毕业要求的对应关系表 课程目标 课程子目标 对应课程内容 对应毕业要求 Understand the 1.1 All experiments principles and experimental skills of the biochemistry lab. 课程目标1 Understand the 1.2 principles and All experiments experimental skills of the biochemistry lab. Understand the principles and 2.1 All experiments experimental skills of the biochemistry lab. 课程目标2 Understand the principles and 2.2 All experiments experimental skills of the biochemistry lab
（课程目标规定某一阶段的学生通过课程学习以后，在发展德、智、体、美、劳等方面 期望实现的程度，它是确定课程内容、教学目标和教学方法的基础。） 课程目标 1：Help students to recapitulate the basic knowledge covered in the course of Biochemistry and Molecular biology. 1.1 Help students to recapitulate the basic knowledge learnt from the theoretical class 1.2 Help students to understand the principles of several basic biochemistry experiments. 课程目标 2：Prepare students for the future work in research labs 2.1 Training student the basic biochemistry experimental skills 2.2 Help students to learn lab report writing skills （要求参照《普通高等学校本科专业类教学质量国家标准》，对应各类专业认证标准， 注意对毕业要求支撑程度强弱的描述，与“课程目标对毕业要求的支撑关系表一致）（五号 宋体） （三）课程目标与毕业要求、课程内容的对应关系 表 1：课程目标与课程内容、毕业要求的对应关系表 课程目标 课程子目标 对应课程内容 对应毕业要求 课程目标 1 1.1 All experiments Understand the principles and experimental skills of the biochemistry lab. 1.2 All experiments Understand the principles and experimental skills of the biochemistry lab. 课程目标 2 2.1 All experiments Understand the principles and experimental skills of the biochemistry lab. 2.2 All experiments Understand the principles and experimental skills of the biochemistry lab
(大类基础课程、专业教学课程及开放选修课程按照本科教学手册中各专业拟定的毕业 要求填写“对应毕业要求”栏。通识教育课程含通识选修课程、新生研讨课程及公共基础课 程，面向专业为工科、师范、医学等有专业认证标准的专业，按照专业认证通用标准填写“对 应毕业要求”栏：面向其他尚未有专业认证标准的专业，按照本科教学手册中各专业拟定的 毕业要求填写“对应毕业要求”栏。) 三、教学内容 (具体描述各章节教学目标、教学内容等。实验课程可按实验模块描述) Experiment 1 Introduction To Basic Technique Of Biochemistry 1.教学目标Aims(五号宋体) (1)To understand the rule of the biochemistry lab. (2)Be familiar with how to write the report. (3)Grasp how to use glass pipettes,centrifuge,spectrophotometry. 2.教学重难点Key points: (1)How to use the glass pipette (2)How to use the centrifuge (3)How to use spectrophotometry 3.教学内容Content: Grasp the basic technique of biochemistry experiment,including centrifuge,the use of the spectrophotometer.Recognize the volume on the glass pipette,and the eyes should be on the same horizontal line with the concave liquid level.Balance the tubes with a scale and put the two tubes in the opposite position.For the spectrophotometer:Check the wavelength before use and open the lid to adjust "0", close the lid to adjust“100%”. 4.教学方法Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental steps to students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation:
（大类基础课程、专业教学课程及开放选修课程按照本科教学手册中各专业拟定的毕业 要求填写“对应毕业要求”栏。通识教育课程含通识选修课程、新生研讨课程及公共基础课 程，面向专业为工科、师范、医学等有专业认证标准的专业，按照专业认证通用标准填写“对 应毕业要求”栏；面向其他尚未有专业认证标准的专业，按照本科教学手册中各专业拟定的 毕业要求填写“对应毕业要求”栏。） 三、教学内容 （具体描述各章节教学目标、教学内容等。实验课程可按实验模块描述） Experiment 1 Introduction To Basic Technique Of Biochemistry 1.教学目标 Aims （五号宋体） （1） To understand the rule of the biochemistry lab. （2） Be familiar with how to write the report. （3） Grasp how to use glass pipettes, centrifuge, spectrophotometry. 2. 教学重难点 Key points: （1） How to use the glass pipette （2） How to use the centrifuge （3） How to use spectrophotometry 3. 教学内容 Content: Grasp the basic technique of biochemistry experiment, including centrifuge, the use of the spectrophotometer. Recognize the volume on the glass pipette, and the eyes should be on the same horizontal line with the concave liquid level. Balance the tubes with a scale and put the two tubes in the opposite position. For the spectrophotometer: Check the wavelength before use and open the lid to adjust “0”, close the lid to adjust “100%”. 4. 教学方法 Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental steps to students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5. 教学评价 Evaluation:
The teacher will evaluate the experimental procedure done by the students on-site.The students will finish the experimental report.Student should summarize the experimental result and attach necessary documents from their lab experiment. The students should also discuss the reason for experimental failures.The teacher will evaluate the report according to their experimental result and discussion. The students need to know the rules to choose and use glass pipette;how to measure the concentration of unknown solution using the standard curve method;how to use a centrifuge Experiment 2 Effects of some factors on enzymatic activity (a-salivary amylase) 1.教学目标Aims: (1)To understand the properties of the enzyme and the effects of some factors such as temperature and pH on enzymatic activity. (2)Be familiar with and deepen the understanding of the chemical nature of enzymes. (3)Grasp the saliva amylase in different temperatures,pH value,under the condition of the vitality of hydrolyzed starch size,and the chloride ion, copper ion on its enzymatic activity. 2.教学重难点Key points: Compare tubes in groups:1,2,3 to determine how the temperature influence enzymatic activity:3,4,5 to understand how pH affects enzymatic activity;3,6,7,8 to understand how different ions affect a-salivary amylase activity. 3.教学内容Content: All enzyme-catalyzed reactions are affected by temperature.The higher the temperature,the higher the reaction rate.However,enzymes are proteins that become denatured at high temperature.So each enzyme has an optimum temperature at which it operates at maximal efficiency.An enzyme's optimum temperature is usually close to the normal temperature of the organism it comes from(around 37C )Changes in pH also affect enzymes through alteration of the tertiary structure of the enzyme or active site.Furthermore,activators and inhibitors can play a key role in regulation of enzymatic activity. 4.教学方法Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental
The teacher will evaluate the experimental procedure done by the students on-site. The students will finish the experimental report. Student should summarize the experimental result and attach necessary documents from their lab experiment. The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. The students need to know the rules to choose and use glass pipette; how to measure the concentration of unknown solution using the standard curve method; how to use a centrifuge Experiment 2 Effects of some factors on enzymatic activity (salivary amylase) 1. 教学目标 Aims: （1） To understand the properties of the enzyme and the effects of some factors such as temperature and pH on enzymatic activity. （2） Be familiar with and deepen the understanding of the chemical nature of enzymes. （3） Grasp the saliva amylase in different temperatures, pH value, under the condition of the vitality of hydrolyzed starch size, and the chloride ion, copper ion on its enzymatic activity. 2.教学重难点 Key points: Compare tubes in groups: 1,2,3 to determine how the temperature influence enzymatic activity; 3,4,5 to understand how pH affects enzymatic activity; 3,6,7,8 to understand how different ions affect a-salivary amylase activity. 3. 教学内容 Content: All enzyme-catalyzed reactions are affected by temperature. The higher the temperature, the higher the reaction rate. However, enzymes are proteins that become denatured at high temperature. So each enzyme has an optimum temperature at which it operates at maximal efficiency. An enzyme’s optimum temperature is usually close to the normal temperature of the organism it comes from(around 37℃ ). Changes in pH also affect enzymes through alteration of the tertiary structure of the enzyme or active site. Furthermore, activators and inhibitors can play a key role in regulation of enzymatic activity. 4.教学方法 Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental
steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation: The teacher will evaluate the experimental procedure done by the students on-site.The students will finish the experimental report.The students should summarize the experimental result and attach necessary documents from their lab experiment.The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 3 The Lowry Assay for Protein 1.教学目标Aims: (1)Understand the basic methods and the principle of the determination of protein content in the samples. (2)Familiar with Folin phenol reagent method principle and the method for determination of protein content. (3)Grasp the principle and method of Folin-phenol reagent to evaluate protein content. 2.教学内容Content: The Lowry method relies on two different reactions.The first is the formation of a copper ion complex with amide bonds,forming reduced copper in alkaline solutions.This is called a "Biuret"chromophore.The second is the reduction of Folin-Ciocalteu reagent (phosphomolybdate and phosphotungstate)by tyrosine and tryptophan residues.The reduced Folin-Ciocalteu reagent is blue and thus detectable with a spectrophotometer in the range of 500-750 nm.The Biuret reaction itself is not all that sensitive.Using the Folin-Ciocalteu reagent to detect reduced copper makes the assay nearly 100 times more sensitive than the Biuret reaction alone. 2.教学重难点Key points: Draw standard curve:Using the protein content of each standard tube as X axis, OD value as Y axis,Draw the standard curve.The standard curve can be used to determine the protein content of specimen. 4.教学方法Teaching method: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental
steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价 Evaluation： The teacher will evaluate the experimental procedure done by the students on-site. The students will finish the experimental report. The students should summarize the experimental result and attach necessary documents from their lab experiment. The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 3 The Lowry Assay for Protein 1. 教学目标 Aims: （1） Understand the basic methods and the principle of the determination of protein content in the samples. （2） Familiar with Folin phenol reagent method principle and the method for determination of protein content. （3） Grasp the principle and method of Folin-phenol reagent to evaluate protein content. 2. 教学内容 Content: The Lowry method relies on two different reactions. The first is the formation of a copper ion complex with amide bonds, forming reduced copper in alkaline solutions. This is called a "Biuret" chromophore. The second is the reduction of Folin-Ciocalteu reagent (phosphomolybdate and phosphotungstate) by tyrosine and tryptophan residues. The reduced Folin-Ciocalteu reagent is blue and thus detectable with a spectrophotometer in the range of 500-750 nm. The Biuret reaction itself is not all that sensitive. Using the Folin-Ciocalteu reagent to detect reduced copper makes the assay nearly 100 times more sensitive than the Biuret reaction alone. 2.教学重难点 Key points: Draw standard curve：Using the protein content of each standard tube as X axis, OD value as Y axis, Draw the standard curve. The standard curve can be used to determine the protein content of specimen. 4.教学方法 Teaching method: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental
steps to the students orally and remind the students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation: (1)The teacher should supervise students during the whole process of experiment and record if the students have attentively and successfully finished the experiment. (2)The students will hand in the experiment report before deadline.In the report, the raw data and the calculation result should be included and the students should summarize the experimental result and attach necessary documents (e.g. standard curve printed and made in excel)from their lab experiment.Student should also discuss the reason for experimental failures.Teacher will evaluate the report according to their experimental result and discussion. Experiment 4 Determination Of Cholesterol In HDL 1.教学目标Aims: Comprehend the method of determination of cholesterol in HDL. Learn about the importance of HDL in vivo and its clinical significance. 2.教学重难点Key points: (1) Understand the clinical significance of HDL cholesterol in blood (2) Less than 40 mg/dL Low level:A major risk factor for CAD (coronary artery disease).60 mg/dL and above High level:considered protective against CAD. 3.教学内容Content: Phosphotungstic acid and magnesium chloride can precipitate CM.VLDL and LDL of the serum,then remove them with centrifugation.Extract cholesterol with anhydrous alcohol and acetic ether (extract solution to precipitate protein,and then centrifuge to remove precipitation.Add development solvent (concentrated sulfuric acid and iron trichloride),which can react with cholesterol and produce stable purple compound.The shade of purple is in direct proportion to the amount of cholesterol.Using the standard cholesterol solution known concentration as control can determine the amount of cholesterol in HDL. 4.教学方法Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental
steps to the students orally and remind the students of the key points that determine the successful accomplishment of the experiment. 5.教学评价 Evaluation: （1） The teacher should supervise students during the whole process of experiment and record if the students have attentively and successfully finished the experiment. （2） The students will hand in the experiment report before deadline. In the report, the raw data and the calculation result should be included and the students should summarize the experimental result and attach necessary documents (e.g. standard curve printed and made in excel) from their lab experiment. Student should also discuss the reason for experimental failures. Teacher will evaluate the report according to their experimental result and discussion. Experiment 4 Determination Of Cholesterol In HDL 1.教学目标 Aims: Comprehend the method of determination of cholesterol in HDL. Learn about the importance of HDL in vivo and its clinical significance. 2.教学重难点 Key points: （1） Understand the clinical significance of HDL cholesterol in blood （2） Less than 40 mg/dL Low level: A major risk factor for CAD (coronary artery disease). 60 mg/dL and above High level: considered protective against CAD. 3. 教学内容 Content: Phosphotungstic acid and magnesium chloride can precipitate CM、VLDL and LDL of the serum, then remove them with centrifugation. Extract cholesterol with anhydrous alcohol and acetic ether（extract solution ）to precipitate protein, and then centrifuge to remove precipitation. Add development solvent (concentrated sulfuric acid and iron trichloride), which can react with cholesterol and produce stable purple compound. The shade of purple is in direct proportion to the amount of cholesterol. Using the standard cholesterol solution known concentration as control can determine the amount of cholesterol in HDL. 4.教学方法 Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental
steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation: (1)The teacher should supervise students during the whole process of experiment and record if the students have attentively and successfully finished the experiment. (2)The students will hand in the experiment report before deadline.In the report,the raw data and the result after calculation should be included and the students should discuss the clinical significance of the result from the experiment.Student should also discuss the reason for experimental failures. Teacher will evaluate the report according to their experimental result and discussion. Experiment 5 Purification of y-globulin From Serum 1.教学目标Aims: (1) Understand the principle of gel filtration, (2) Understand the protein purification method (3)Master gel filtration procedure. 2.教学重难点Key Point: (1) Principle of salting out method for protein purification (2) Principle of gel filtration (3) Packing gel filtration column with Sephorax G25 (4)Protein and NH4S04 detection by ninhydrin solution and Nessler solution. 3.教学内容Content: Separating y-globulin from serum using salting out method using NH,SO,solution. Purify y-globulin by gel filtration and detecting separated protein and NH SO,using ninhydrin solution and Nessler solution.The students will learn to perform salting out and to pack gel filtration column themselves. 4.教学方法Teaching Methods:
steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价 Evaluation: （1） The teacher should supervise students during the whole process of experiment and record if the students have attentively and successfully finished the experiment. （2） The students will hand in the experiment report before deadline. In the report, the raw data and the result after calculation should be included and the students should discuss the clinical significance of the result from the experiment. Student should also discuss the reason for experimental failures. Teacher will evaluate the report according to their experimental result and discussion. Experiment 5 Purification of -globulin From Serum 1. 教学目标 Aims: （1） Understand the principle of gel filtration, （2） Understand the protein purification method （3） Master gel filtration procedure. 2. 教学重难点 Key Point: （1） Principle of salting out method for protein purification （2） Principle of gel filtration （3） Packing gel filtration column with Sephorax G25 （4） Protein and NH4SO4 detection by ninhydrin solution and Nessler solution. 3. 教学内容 Content: Separating -globulin from serum using salting out method using NH4SO4 solution. Purify -globulin by gel filtration and detecting separated protein and NH4SO4 using ninhydrin solution and Nessler solution. The students will learn to perform salting out and to pack gel filtration column themselves. 4.教学方法 Teaching Methods:
The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation: The teacher will evaluate the experimental procedure done by the students on-site.The students will finish the experimental report.The students should summarize the experimental result and attach necessary documents from their lab experiment.The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 6 Quantitative Determination Of The Serum Protein With Cellulose Acetate Membrane Electrophoresis 4.教学目标Aims: (1)Comprehend the method of electrophoresis. (2) Familiar with the percentage values of human serum protein. (3)Master the method of cellulose acetate membrane electrophoresis to separate and identify serum protein. 2.教学重难点Key points: Principles to separate serum proteins with cellulose acetate membrane electrophoresis 3.教学内容Content: The isoelectric point (PI)of various kinds of serum proteins are generally lower than 7.0,in alkaline buffer solution(PH 8.6),serum proteins are negatively charged, they will migrate toward anode in an electric field.The rate of migration will depend on the strength of their net surface charges,shape and size of the proteins.Because the PI of the different serum proteins are different,so their net surface charges are different.Furthermore,the size shape are distinct,consequently,the rate of migration is different too. 4.教学方法Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental
The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价 Evaluation： The teacher will evaluate the experimental procedure done by the students on-site. The students will finish the experimental report. The students should summarize the experimental result and attach necessary documents from their lab experiment. The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 6 Quantitative Determination Of The Serum Protein With Cellulose Acetate Membrane Electrophoresis 4. 教学目标 Aims: （1） Comprehend the method of electrophoresis. （2） Familiar with the percentage values of human serum protein. （3） Master the method of cellulose acetate membrane electrophoresis to separate and identify serum protein. 2.教学重难点 Key points: Principles to separate serum proteins with cellulose acetate membrane electrophoresis 3.教学内容 Content: The isoelectric point (PI) of various kinds of serum proteins are generally lower than 7.0, in alkaline buffer solution (PH 8.6), serum proteins are negatively charged, they will migrate toward anode in an electric field. The rate of migration will depend on the strength of their net surface charges、shape and size of the proteins. Because the PI of the different serum proteins are different, so their net surface charges are different. Furthermore, the size 、shape are distinct, consequently, the rate of migration is different too. 4.教学方法 Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental
steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation: The teacher will evaluate the experimental procedure done by the students on-site.The students will finish the experimental report.The students should summarize the experimental result and attach necessary documents from their lab experiment.The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 7 Isolation and purification of nucleic acid 教学目标Aims: (1) Understand the distribution of DNA and RNA in the cell. (2) Master the principle and method of isolation and purification of nuclei from rat liver cells. 2.教学重难点Key points: Principle and method to separate cell nuclei 3.教学内容Content: The method that is commonly used for isolation and purification of nuclei involves the mechanical disruption and fractionation of cells into their different subcellular components (e.g.nucleus,microsomes,mitochondria,cytoplasm).A very common mean of fractionating cells takes advantage of the fact that different components of a cell have different sedimentation coefficients.The factors that determine the sedimentation coefficient for different components are the radius and the effective density of these microscopic objects.The sedimentation coefficient is different for all types of subcellular structures,such as nuclei,mitochondria, microsomes (small pieces of disrupted ER),lysosomes and ribosomes.Therefore,we can isolate and purify such subcellular components from each other merely by centrifuging a mixture of them (a crude cell lysate)at different speeds.For example, at a speed that produces a force 600 times the earth's gravity,the nuclei of cells will pellet at the bottom of a tube,while most of the other cellular particles will remain in suspension.At a centrifuge speed that produces a force 10000 times that of gravity,large organelles such as mitochondria and chloroplasts will usually pellet.Much higher levels of force (100000 times gravity)are usually needed to
steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价 Evaluation： The teacher will evaluate the experimental procedure done by the students on-site. The students will finish the experimental report. The students should summarize the experimental result and attach necessary documents from their lab experiment. The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 7 Isolation and purification of nucleic acid 教学目标 Aims: （1） Understand the distribution of DNA and RNA in the cell. （2） Master the principle and method of isolation and purification of nuclei from rat liver cells. 2.教学重难点 Key points: Principle and method to separate cell nuclei 3.教学内容 Content: The method that is commonly used for isolation and purification of nuclei involves the mechanical disruption and fractionation of cells into their different subcellular components (e.g. nucleus, microsomes, mitochondria, cytoplasm). A very common mean of fractionating cells takes advantage of the fact that different components of a cell have different sedimentation coefficients. The factors that determine the sedimentation coefficient for different components are the radius and the effective density of these microscopic objects. The sedimentation coefficient is different for all types of subcellular structures, such as nuclei, mitochondria, microsomes (small pieces of disrupted ER), lysosomes and ribosomes. Therefore, we can isolate and purify such subcellular components from each other merely by centrifuging a mixture of them (a crude cell lysate) at different speeds. For example, at a speed that produces a force 600 times the earth’s gravity, the nuclei of cells will pellet at the bottom of a tube, while most of the other cellular particles will remain in suspension. At a centrifuge speed that produces a force 10000 times that of gravity, large organelles such as mitochondria and chloroplasts will usually pellet. Much higher levels of force (> 100000 times gravity) are usually needed to
pellet microsomes and ribosomes. 4.教学方法Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class.The teacher should display the basic experimental steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价Evaluation: The teacher will evaluate the experimental procedure done by the students on-site.The students will finish the experimental report.The students should summarize the experimental result and attach necessary documents from their lab experiment.The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 8 Assaying of nucleic acid 1.教学目标Aims: (1) Comprehend the principle and method of mensuration of DNA and RNA. (2) Familiar with the principle and method of determination of DNA and RNA content. 2.教学重难点Key points: (1) Principle and method to extract DNA and RNA (2) Principle and method to quantify DNA and RNA 3.教学内容Content: There are two types of nucleic acid:DNA and RNA.DNA chiefly resides in cellular nucleus,and in cytoplasm RNA is most plentiful.In alkaline solution RNA cab be hydrolyzed,its ribose dehydrated to form furfurol,then furfurol can Kondens with orcinol to synthesize a green compound,the shade of green is in direct proportion to the amount of RNA.Heated in perchloric acid solution,DNA can be hydrolysed, its ribodesose dehydrated to form w-hydroxy-keto-pentanal,then it can react with diphenylamine to produce a blue compound,which chemical property is not clear,but the shade of blue is in direct proportion to the amount of DNA. 4.教学方法Teaching Methods:
pellet microsomes and ribosomes. 4.教学方法 Teaching Methods: The teacher should recapitulate the key concept of the corresponding chapter taught in the theoretical class. The teacher should display the basic experimental steps to the students orally and remind students of the key points that determine the successful accomplishment of the experiment. 5.教学评价 Evaluation： The teacher will evaluate the experimental procedure done by the students on-site. The students will finish the experimental report. The students should summarize the experimental result and attach necessary documents from their lab experiment. The students should also discuss the reason for experimental failures. The teacher will evaluate the report according to their experimental result and discussion. Experiment 8 Assaying of nucleic acid 1.教学目标 Aims: （1） Comprehend the principle and method of mensuration of DNA and RNA. （2） Familiar with the principle and method of determination of DNA and RNA content. 2.教学重难点 Key points: （1） Principle and method to extract DNA and RNA （2） Principle and method to quantify DNA and RNA 3.教学内容 Content: There are two types of nucleic acid: DNA and RNA. DNA chiefly resides in cellular nucleus, and in cytoplasm RNA is most plentiful. In alkaline solution RNA cab be hydrolyzed, its ribose dehydrated to form furfurol, then furfurol can Kondens with orcinol to synthesize a green compound, the shade of green is in direct proportion to the amount of RNA. Heated in perchloric acid solution, DNA can be hydrolysed, its ribodesose dehydrated to form ω-hydroxy-keto-pentanal, then it can react with diphenylamine to produce a blue compound, which chemical property is not clear, but the shade of blue is in direct proportion to the amount of DNA. 4.教学方法 Teaching Methods:
