Laboratory III
Code: 100978 ECTS Credits: 3| Degree | Type | Year | Semester |
|---|---|---|---|
| 2500502 Microbiology | OB | 2 | 1 |
Contact
- Name:
- Susana Campoy Sanchez
- Email:
- susana.campoy@uab.cat
Teaching groups languages
You can check it through this link. To consult the language you will need to enter the CODE of the subject. Please note that this information is provisional until 30 November 2023.
Teachers
- Ignacio Gerardo Alvarez Perez
- Maria Area Navarro
- Susana Campoy Sanchez
- Alicia Roque Cordova
Prerequisites
It is desirable to have coursed or are coursing first semester of the second year of the Degree of Microbiology and the previous integrated laboratory I and II courses.
It is mandatory that the student has passed the safety and biosafety tests that will be found in the Virtual Campus. It is also compulsory to know and accept the operating rules of the teaching laboratories of the Faculty of Biosciences and accomplish all the specific instructions of the teachers.
Objectives and Contextualisation
This course introduces students to basic molecular and immunological techniques for experimentation in a Molecular Biology laboratory.
The specific objectives to be achieved are defined in the following points:
• To know how to apply biochemical fractionation and other analysis techniques.
• Perform nucleic acid extraction and electrophoresis techniques.
• To know how to use basic recombinant DNA techniques such as restriction endonucleases, DNA cloning in vectors, DNA transformation, etc.
• To know the methodologies for the labeling of DNA probes and to apply specific detection techniques of these labeled probes by DNA-DNA hybridization.
• Use and know how to apply PCR techniques and/or quantitative PCR.
• To know how to use immunological techniques of precipitation and agglutination or ELISA and understand their application in the identification and typing of microorganisms.
• Integrate knowledge of molecular biology, biochemistry, microbiology and immunology for the cloning, overexpression, purification, and detection of a bacterial protein.
Competences
- Adapt to new situations.
- Apply knowledge of theory to practice
- Communicate orally and in writing.
- Design and apply methods and strategies for isolating and selecting new microorganisms and for genetically manipulating microorganisms of interest.
- Develop creativity and initiative.
- Display a capacity for analysis, synthesis, organisation, planning and decision-making.
- Use molecular and immunological techniques in the characterisation of microorganisms and materials of biological origin.
- Work individually or in groups, in multidisciplinary teams and in an international context.
Learning Outcomes
- Adapt to new situations.
- Apply electrophoretic techniques for separating DNA and proteins.
- Apply knowledge of theory to practice
- Apply methodologies for detecting and analysing different metabolites in biological preparations.
- Apply procedures based on the use of specific probes and on hybridisation with nucleic acids.
- Apply procedures for purifying gene products produced by microorganisms.
- Apply suitable methods for analysing enzyme activity.
- Communicate orally and in writing.
- Describe and apply procedures and techniques for cloning DNA.
- Describe and apply techniques for extracting whole DNA, chromosomal DNA, plasmidic DNA and virus DNA.
- Describe and use immunological techniques.
- Describe and use techniques for amplifying DNA.
- Develop creativity and initiative.
- Display a capacity for analysis, synthesis, organisation, planning and decision-making.
- Interpret the results obtained when using different molecular techniques.
- Process tissues and cells to obtain sub-cellular fractions and characterise these biochemically.
- Use enzymes of molecular biology to modify DNA and describe the procedures.
- Use microbial vectors for the expression of genes of interest in microorganisms.
- Use the advanced instruments and methodologies of a biochemistry laboratory.
- Use the basic techniques for handling and analysing proteins and nucleic acids
- Work individually or in groups, in multidisciplinary teams and in an international context.
Content
The subject will be organized in two distinct parts:
• Monographic methodological sessions lasting approximately three and four hours depending on each session and scheduled weekly in 3 groups of 3 or 4 sessions depending on each module.
• Integrated practice, lasting 19-hours that will be intensive for one week
Monographic sessions
MODULE 1
Sessions 1, 2 and 3. Cell fractionation techniques and biochemical analysis. Characterization of the fractions by enzymatic markers. Techniques of cold labeling probes, of dot blot and hybridization.
MODULE 2
Sessions 4 and 5. Nucleic acid and restriction techniques.
Session 6. PCR techniques.
Session 7. Techniques for detecting interactions between DNA and protein.
MODULE-3
Sessions 8 to 11. Precipitation of immunoglobulins with ammonium sulfate. Determination of the concentration of antibodies in serum by ELISA. Separation of splenocytes by density gradient. Determination of phagocytosis by a monocyte cell line using the flow cytometry technique.
INTEGRATED PRACTICE
Sessions 1 and 2. Cloning of a bacterial gene and overexpression of the gene product.
Session 3. Purification and quantification of the protein.
Sessions 4 and 5. Detection of the protein by Western blot.
Methodology
The subject will be taught in small groups of students.
To acquire the specific competencies of the subject attendance to the practical sessions is mandatory. If a student for justified and unforeseeable reasons does not attend a practice session(s), he/she must communicate to the responsible teacher and present the corresponding certificate as soon as possible to the Degree coordinator. It is understood for justified cause those enclosed in the Faculty of Biosciences evaluation criteria (section 1).
The students will have a manual of the course before the beginning of the practical sessions. The student must bring his/her gown, laboratory glasses, a notebook, and the manual (available in the Virtual Campus or where indicated by the teaching staff).
The students will work in small groups. At the beginning of each session, the teacher will make a brief theoretical explanation of the content of the practice and the experiences to be carried out.
To achieve a good performance and acquire the competencies corresponding to this course, the student must make a comprehensive reading of the practice manual, familiarizing with the practices that will be carried out in each session as well as with the methodology that should apply in each case.
Annotation: Within the schedule set by the centre or degree programme, 15 minutes of one class will be reserved for students to evaluate their lecturers and their courses or modules through questionnaires.
Activities
| Title | Hours | ECTS | Learning Outcomes |
|---|---|---|---|
| Type: Directed | |||
| Classroom sessions | 2 | 0.08 | 1, 3, 7, 4, 5, 2, 10, 12, 11, 13, 15, 16, 8, 21, 19, 14, 17, 20 |
| Laboratory sessions | 53 | 2.12 | 1, 3, 7, 4, 5, 6, 2, 9, 10, 12, 11, 13, 18, 15, 16, 8, 21, 19, 14, 17, 20 |
| Type: Supervised | |||
| Tutorship | 1 | 0.04 | 1, 3, 7, 4, 5, 6, 2, 9, 10, 12, 11, 13, 18, 15, 16, 8, 21, 19, 14, 17, 20 |
| Type: Autonomous | |||
| Comprehensive reading of the practice manual | 4 | 0.16 | 1, 3, 7, 4, 5, 6, 2, 9, 10, 12, 11, 13, 18, 15, 16, 8, 21, 19, 14, 17, 20 |
| Study | 4 | 0.16 | 1, 3, 10, 12, 11, 13, 15, 21, 14 |
| Writing reports or questionnaires | 6.5 | 0.26 | 3, 7, 4, 5, 6, 2, 9, 12, 18, 15, 16, 8, 19, 17 |
Assessment
The evaluation of the subject is continuous and distributed in the different sections:
MODULE 1:
The assessment is through a questionnaire referred to sessions 1, 2, and 3. This questionnaire must be submitted when indicated by the teaching staff. To pass this module the qualification obtained in the questionnaire must be equal to or greater than 5.
MODULE 2:
A questionnaire is made at the end of the practical sessions. The questionnaire allows for assessing if the student has reached the competencies associated with the module. To pass this module the obtained score in the questionnaire must be equal to or greater than 5.
MODULE 3:
This module is assessed by conducting a questionnaire at the end of session 12. To pass this module the score obtained in the questionnaire must be equal to or greater than 5.
INTEGRATED PRACTICE:
The assessment of integrated practice is composed of two distinct aspects:
A) 8 points out of 10 come from the qualification obtained in a questionnaire.
B) 2 points out of 10 are obtained by the achievement in the laboratory of the objectives set in each session.
To pass this module, the final qualification must be equal to or greater than 5.
PRIOR KNOWLEDGE OF ACTIVITIES TO PRACTICAL SESSIONS:
This section takes into account the prior knowledge of the students before beginning the practical session that derives from the previous comprehensive reading of the practice manual.
The assessment is done using random questionnaires delivered at the beginning of some sessions. The final grade of this section is the average grade of the different tests carried out.
The final grade of the subject is the weighted average of the grades obtained in each of the different sections taking into account that each module will represent 20% of the final grade of the subject and the integrated practice will correspondto 30%. The remaining 10% depends on the qualification obtained in the previous knowledge section.
Note that it is possible to penalize with a maximum of 40% with respect to 100% of the final qualification of the subject incorrect attitudes of the student in the laboratory such as lack of punctuality, not performing autonomous work indicated by the teacher, non-respectful behavior with colleagues or teachers, improper use of laboratory equipment and equipment, etc.
To pass the course you must obtain a final grade equal to or greater than 5, being necessary to overcome each of the modules and the integrated practice.
Students who do not pass the evaluation of the different modules or the integrated practice should perform the scheduled retake examination at the end of the semester, opting for a maximum score of 8 points on the 10 possible of recovered module(s).
Students who do not obtain the minimum qualification required to pass each of the integrated laboratory modules will not pass the subject. In this case, the maximum final grade for the subject will be 4.
In the second enrollment, the students can only perform the non-passed modules. The students should contact the coordinator of their pending module(s). In the case they pass the module(s) in this new enrollment, the final grade of the course will be the weighted average of the grades of the passed module(s) in this academic year with those of passed module(s) in previous registrations. In case of not overcoming the module(s), the maximum final grade of the subject will be 4.
Given that attendance at the scheduled activities is mandatory, absence from any of them must be justified and communicated to the person in charge of the subject and to the coordinator of the Degree as defined in the Methodology section. To pass the subject, a global attendance of at least 80% of the scheduled sessions is required, as well as obtaining the minimum mark set for each module. Therefore, it is considered that a student obtains the qualification of "not evaluable" when he has attended less than 80% of the scheduled sessions.
SINGLE ASSESSMENT:
The students who take advantage of the single evaluation must attend the laboratory sessions scheduled in the degree calendar since these are compulsory. The single evaluation consists of a test, which will be divided into four questionnaires on the contents covered in modules 1, 2, 3, and the integrated practice. Each of the questionnaires will have a score of 10 points and it will be necessary to obtain a grade equal to or greater than 5 in each of them to pass it and therefore pass the subject. The final mark of this will be the weighted average of the four parts of the test, where the mark of the questionnaires of modules 1, 2, and 3 will have a relative weight of 22% each, while the part associated with the integrated practice will correspond to 34% of the total subject. In case of not passing any of the modules, a recovery evaluation will be scheduled under the same conditions as the former. It will be necessary to pass each module separately in order to average the qualification, which in all cases must be equal to or greater than 5.
Assessment Activities
| Title | Weighting | Hours | ECTS | Learning Outcomes |
|---|---|---|---|---|
| Integrated practice's test type questionnaire | 24% | 1.5 | 0.06 | 1, 3, 5, 6, 2, 9, 12, 11, 13, 18, 15, 8, 21, 19, 14, 17, 20 |
| Laboratory work performance during the Integrated practice | 6% | 0 | 0 | 1, 3, 7, 5, 6, 2, 9, 10, 12, 11, 18, 15, 19, 17, 20 |
| Prior knowledge of activities to practical sessions | 10% | 0 | 0 | 3, 8, 14 |
| Questionnaire of Module 1 | 20% | 0 | 0 | 1, 3, 7, 4, 13, 15, 16, 8, 21, 19, 14, 20 |
| Test type questionnaire of Module 2 | 20% | 2 | 0.08 | 1, 3, 2, 9, 10, 12, 13, 15, 8, 21, 14, 17, 20 |
| Test type questionnaire of Module 3 | 20% | 1 | 0.04 | 1, 3, 5, 11, 13, 15, 8, 21, 14 |
Bibliography
Students will have a practice manual of the course that details the objectives of each session along with the protocols that will be used. Also in the dossier, will be recorded the bibliography associated to each one of the different sessions.
Software
The usual Microsoft environment programs will be used.