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Integrated Laboratory Class 4

Code: 100925 ECTS Credits: 3
2024/2025
Degree Type Year
2500253 Biotechnology OB 2

Contact

Name:
Antonio Javier Moral Vico
Email:
antoniojavier.moral@uab.cat

Teachers

Octavi Marti Sistac
Carles Barril Basil
Jesus Aranda Rodriguez

Teaching groups languages

You can view this information at the end of this document.


Prerequisites

It is recommended to attend simultaneously, or have taken, the theory subjects corresponding to the contents of the laboratory practices.

You must have passed the safety test in laboratories. The test is answered in the corresponding space of the Virtual Campus and the information that must be consulted is in the space for communication of the Degree in Biotechnology.


Objectives and Contextualisation

The Integrated Laboratory 4 is the fourth subject of a set of 6 that are distributed over the 6 semesters corresponding to the first three years of the Degree in Biotechnology.

The training objectives of these subjects focus on the acquisition of competences within the framework of the practical training of the student.

The contents are organized in a growing order of complexity and associated to the needs and progress of the theoretical contents of the Degree.

The Integrated Laboratory 4 has as its training objectives the acquisition of practical skills in 4 modules:

- Animal Physiology

- Molecular Microbiology

- Bioreactors

- Numerical Methods and Computer Applications.

These modules are grouped into two blocks:

1- Animal Physiology and the molecular biology of microoorganisms.

2- Fundamentals for the design of bioreactors and development of bioprocesses: The aim is to acquire basic knowledge in the design, operation and characterization of the main types of bioreactors, and the approach and resolution of the mathematical equations derived from them.


Learning Outcomes

  1. CM22 (Competence) Prioritise the instrumentation necessary for the different techniques for the separation and characterisation of biomolecules.
  2. CM22 (Competence) Prioritise the instrumentation necessary for the different techniques for the separation and characterisation of biomolecules.
  3. CM24 (Competence) Review the general safety standards of a biotechnology laboratory.
  4. KM22 (Knowledge) Relate the physiology of different organs and the various metabolic states of an organism.
  5. KM23 (Knowledge) Recognise the key microscopic characteristics that distinguish prokaryotic cells from eukaryotic cells, and animal cells from plant cells.
  6. SM20 (Skill) Use the basic techniques of manipulation, separation, detection and analysis of proteins and nucleic acids.
  7. SM20 (Skill) Use the basic techniques of manipulation, separation, detection and analysis of proteins and nucleic acids.
  8. SM21 (Skill) Use prokaryotic and eukaryotic cell culture techniques and techniques for the manipulation of biological systems.
  9. SM21 (Skill) Use prokaryotic and eukaryotic cell culture techniques and techniques for the manipulation of biological systems.
  10. SM22 (Skill) Use analytical methodologies for the assay of biological activity of cellular components.
  11. SM22 (Skill) Use analytical methodologies for the assay of biological activity of cellular components.

Content

The subject is structured in 4 modules.

Animal Physiology

Contents: they are organized in 4 sessions of 3 hours that are done in the laboratory (1 and 2) or computer room (3 and 4).

1. Cardiovascular and respiratory adaptation to physical exercise. Influence of sex and hydrostatic pressure.

Determination of heart rate, systolic and diastolic blood pressure, and saturation of blood oxygen in various conditions before and after exercise. Empirical verification of the effect of hydrostatic pressure on blood pressure.

2. Study of the human electrocardiogram (ECG). Cardiac respiratory physiological arrhythmia (ACRF).

Acquisition of the ECG and identification of the different waves that comprise it. ECG and ACRF.

3. Identification of structures and gene expression in the central nervous system: virtual brain stereotactic brain atlas.

Physiological neuroanatomy study of the brain of the mouse to identify some of the main structures  that it contains, as well as the degree of gene expression of genes of interest. An interactive 3D steroidal brain mouse atlas will be used, with online access to gene expression databases.

4. Statistical analysis of the data obtained for the whole group of practices.

Preliminary inspection of the data obtained in sessions 1 and 2, and subsequent statistical analysis of the results obtained for the entire group of practices. Assessment of the possible statistically significant differences in the results.

Molecular Microbiology

The Molecular Microbiology module is organized into 5 sessions. The practices in these sessions will allow the student to learn the basic techniques of DNA transfer in bacteria, the mechanisms of directed and random mutagenesis used for the genetic modification of prokaryotes, and the mechanisms that allow the study of gene expression and its regulation in bacteria. All these contents will be grouped into the 4 practices that are listed below.

Practice 1 (4h) Transfer of genetic material into prokaryotes

Different methodologies will be used for the incorporation of exogenous DNA into bacteria, such as transformation mechanisms, biparental conjugation, triparental conjugation, and transduction of markers between bacteria.

Practice 2 (2h). Processes of mutagenesis and recombination to obtain new strains

Basic processes for the interchange of bacterial genetic material will be applied, such as experiments to obtain spontaneous mutants, directed mutagenesis, or the integration and / or replacement of genetic material by recombination.

Practice 3 (4h). Use of mobile genetic elements to obtain mutants

Methodologies based on the use of mobile genetic elements for bacterial genetic manipulation will be used. The type ofjumps of these elements will be described, as well as their frequency of movement.

Practice 4 (2h). Control of gene expression in prokaryotes

The tools for the quantification of the bacterial gene expression will be applied, and these methodologies will be used to study regulated promoters as well as toidentify the mechanisms that control their gene expression.

Bioreactors

The practices are organized in 4 sessions of 3 h.

Practice 1 (3h) + Practice 2 (3h). Continuous Stirred Tank Reactor (RCTA)

The operation and the main characteristics of an RCTA type bioreactor are learned. The kinetics of growth of a yeast strain are determined. The stimulus-response techniques are used to determine the distribution of the residence time of the bioreactor, and analyze its hydrodynamic behavior, in particular the mixing characteristics. All this knowledge is included in the equations for the design of RCTA-type bioreactors.

Practice 3 (3h) + Practice 4 (3h). Air-lift reactor.

The operating bases of an Air-lift bioreactor are learned, as well as the different elements involved in its design. The experimental techniques to determine the coefficient of oxygen transfer between a gasphase and a liquid, kLa are used. The influence of the operating conditions of the bioreactor on the properties of gas-liquid transfrence is studied.

The methodology is analyzed to determine the oxygen consumption of a yeast culture.

Numerical Methods and Computer Applications

They are organized in 5 sessions of two and a half hours that are done in the computer room.

Practice 1 (2.5h) Introduction.

The objective is that the student becomes familiar with the programming environment that will be used in these practices. You will see the basic instructions and instructions for the programming of algorithms.

Practice 2 (2.5h) Errors.

The purpose of this practice is to know the limitations of numerical errors. We will see how to detect and control different sources of error in the scientific calculation.

Practice 3 (2.5h) Function Zeros.

In this practice, different numerical methods will be implemented for the calculation of zeros of functions. Its applicability will be studied in different cases.

Practice 4 (2.5h) Integration.

In this practice, polynomial interpolation algorithms will be developed and different numerical methods will be implemented to evaluate defined integrals.

Practice 5 (2.5h) Differential equations.

The objective of this practice is to implement some basic numerical resolution methods for simple cases. You will also see how to use software routines based on more advanced methods.


Activities and Methodology

Title Hours ECTS Learning Outcomes
Type: Directed      
Laboratory session 54.5 2.18 CM22, CM24, KM22, KM23, SM20, SM21, SM22
Type: Autonomous      
Reading protocols 6.5 0.26

The attendance to the classes of this subject is obligatory since they imply an acquisition of competences based on the practical work.

Animal Physiology, Molecular Microbiology and Bioreactors:

The students carry out the experimental work in groups of 2 and under the supervision of the responsible professor.
The practical protocols and, if applicable, the questionnaires for response, will be available on the Virtual Campus of the subject
Before beginning a practical session the student must have read the protocol and know therefore the objectives of the practice, the foundations and the procedures that must be carried out.
If so, you must know the specific safety and waste treatment measures.
When completing the practice of the module of Bioreactors, the students will have to work with the obtained data and present the corresponding reports.

In the practical sessions you must have:
- Protocol and, if applicable, the questionnaire.
- A notebook to collect the information of the experimental work.
- Laboratory baton.
- Safety glasses.
- Permanent marker


Numerical Methods and Computer Applications:

Practical classes in the computer rooms of the faculty.
The students will carry out the proposed work in the practice script under the supervision and direction of the responsible professor. In each session the student will complete a questionnaire on the different problems resolved in practice.
The practical scripts will be available on the Virtual Campus of the subject.
Before beginning a practical session the student must have read the script and know therefore the objectives of the practice and the foundations of the numerical methods that he will have to use.

In the practical sessions you must have:
- The script of the practice.

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.


Assessment

Continous Assessment Activities

Title Weighting Hours ECTS Learning Outcomes
Animal Physiology:Individual evaluation 1.0 0 0 CM24, KM22
Animal physiology: Individual evaluation 1.5 0 0 CM24, KM22
Bioreactors report 1.11 11 0.44
Continuous evaluation Bioreactors 1.11 0 0 CM22, CM24, SM21, SM22
Continuous evaluation Molecular Microbiology 0,67 0 0 CM22, CM24, KM22, KM23, SM20, SM21, SM22
Exam Numeric Methods and Computer Applications 1.89 1 0.04
Questionnaire Molecular Microbiology 1.55 1 0.04 KM23
Questionnaire Numerical Methods and Computer Applications 0.33 1 0.04

Attendance at practical sessions is mandatory. The students will obtain the "Non-Avaluable" qualification when the absence exceeds 20% of the programmed sessions.


Students who do not obtain the minimum qualification of 4, required to be able to pass each one of the modules of the integrated laboratory, will not pass the subject. In this case, the final maximum grade of the subject will be 4.

Due to the fact that the Integrated Laboratory is differentiated in modules, from the second enrollment, repeat students will only have to evaluate the specific modules that have not been exceeded. This exemption will be maintained for a period of three additional tuition fees, participating in a number of assessment activities that can not be granted, at best, the qualification of approved.

The evaluation of each module will be carried out independently, following the criteria detailed below.


The final evaluation of the subject will be obtained from the average assessment of the different modules (33.34%, 22.22%, 22.22%, 22.22%).

 

Animal Physiology

The assessment will consist of two parts:

1.- Evaluation of group work: The group must submit a report on one of the practices carried out, in a format that is comparable to that of a scientific paper, in accordance with the guide that will be provided by the responsible professor. Contribution to the mark of the module: 50%. Skills: E3, E16, T1, T2, T5, T11, T12

2.- Individual/in pairs assessment: Each student must submit the reduced version of a report on different aspects of the practices carried out, according to the teacher's instructions. Contribution to the mark of the module: 50%. Skills: E3, T1, T2, T5, T11, T12


Molecular Microbiology
Two different aspects will be taken into account, on the one hand, the mark obtained in a questionnaire that will be done at the end of the session 5 and which will refer to all the practices that make up this module, and on the other, the achievement of the Targets set in each of the programmed practices. The questionnaire will represent 70% of the final grade of the module while the remaining 30% will depend on the evaluation of the results obtained and the experimental work performed.


Bioreactors
Several different aspects will be taken into account, the quality of the work in the laboratory (20%), the experimental data obtained (10%), questions and problems proposed (20%) and the preparation of the report of the practices (50%). The reports must be submitted before a specific date, which will be announced at the beginning of the laboratory. The delay not justified in the presentation of the reports will imply a punishment in the punctuation of the same.


Numerical Methods and Computer Applications
This module will be evaluated by means of an individual examination at the end of the semester, which will mean 70% of the final mark, and the questionnaires that must be submitted at the end of each practice and which will represent the remaining 30%. In the final exam thestudent will have to solve someproblems similar to those that have been dealt with in the practices.

 

This subject/module does not include the single evaluation system.


Bibliography

Animal Physiology

· Barrett KE, Barman SM, Brooks HL, Yuan JX-J. Ganong. Fisiología Médica. McGraw-Hill Interamericana de España SL, 26a ed, 2020.
· Fox SI. Fisiología Humana. McGraw-Hill Educación, 14a ed, 2017.
· Hall JE, Hall ME. Guyton y Hall. Tratado de Fisiología Médica. Elsevier, 14a ed, 2021.
· Koeppen BM, Stanton BA. Berne & Levy Physiology. Elsevier, 7a ed, 2017.
· Pocock G, Richards CD, Richards DA. Human Physiology. Oxford University Press, 5a ed, 2017.
· Silbernagl S, Despopoulos A. Fisiología. Texto y Atlas. Editorial Médica Panamericana, 7a ed, 2009.
· Tortora GJ, Derrickson BH. Principles of Anatomy and Physiology. Médica Panamericana, 15a ed, 2021.
· Tresguerres J.A.F. Fisiología Humana. McGraw-Hill Interamericana de España SL, 4a ed, 2010.
· Widmaier EP, Raff H, Strang KT. Vander's Human Physiology. The Mechanisms of Body Function. McGraw-Hill Education, 15a ed, 2018.
 
Molecular Microbiology
 
· Erill I, Campoy S, Barbé J. Aeons of distress: an evolutionary perspective on the bacterial SOS response. FEMS Microbiol Rev. 200731(6):637-56.
· Goryshin IY, Jendrisak J, Hoffman LM, Meis R, Reznikoff WS. Insertional transposon mutagenesis by electroporation of released Tn5 transposition complexes.Nat Biotechnol. 2000. 18:97-100.
· Griffiths, A. J. F , Gelbart,WM,Lewontin, R. An Introduction To Genetic Analysis. 2004. 8th ed.
· Hayes F. Transposon-based strategies for microbial functional genomics and proteomics. Annu Rev Genet. 2003. 37:3-29.
· Hoffman LM, Jendrisak JJ, Meis RJ, Goryshin IY, Reznikof SW. Transposome insertional mutagenesis and direct sequencing of microbial genomes. Genetica. 2000. 108:19-24.
· Judson N, Mekalanos JJ. Transposon-based approaches to identify essential bacterial genes.Trends Microbiol. 2000. 8:521-526.
· Kirby JR. In vivo mutagenesis using EZ-Tn5.Methods Enzymol. 2007. 421:17-21.
· Little JW. Mechanism of specific LexA cleavage: autodigestion and the role of RecA coprotease. Biochimie 1991. 73: 411 421.
· Sassanfar M Roberts JW. Nature of the SOS-inducing signal in Escherichia coli. The involvement of DNA replication . J Mol Biol. 1990. 21:79 96.
· Snyde rL. and ChampnessW. Molecular Genetics of Bacteria (3rd)ASM press 2007.
· Voelker LL, Dybvig K.1998.Transposon mutagenesis.Methods Mol Biol. 1998. 104:235-238.
 
Bioreactors
 
· J.C. Merchuck, M.H. Siegel (1988). “Airlift bioreactors in chemical and biological technology”. J. Chem. Tech. Biotecnol. 41, 105-120.
· J.E. Bailey, i D.F. Ollis (1986). Biochemical Engineering Fundamentals. 2nd ed, McGraw Hill Book Company, New York.
· K. Van’t Riet, i H. Tramper (1991). Basic Bioreactor Design. Marcel Dekker Inc., Nova York, USA.
· M.Y. Chisti (1989). Airlift bioreactors. Elsevier Applied Science, Londres.
· O. Levenspiel. Chemical reactor engineering. 3rd ed, Wiley International Ed., New York (2004).
 
 Numerical Methods
 
· A. Aubanell, A. Benseny i A. Delshams, Eines bàsiques del Càlcul numèric, Manuals de la UAB, (1992)
· C. Bonet i altres, Introducció al Càlcul Numèric, Universitat Politècnica de Catalunya, (1989)
· A. Ralston and P. Rabinowitz, A first course in numerical analysis,  McGraw-Hill, 1988.
· L. Elden, L. Wittmeyer-Koch, & H. B.  Nielsen, (2004). Introduction to Numerical Computation. https://users.mai.liu.se/larel04/NumCompEWN2004.p
 

Software

Animal Phisyology

MS Office, BSL-Biopac Student Lab, BrainExplorer, vassarstats.net.  

Molecular Microbiology

It does not apply

Bioreactors

It does not apply

Numerical Methods

Matlab/Octave

 


Language list

Name Group Language Semester Turn
(PLAB) Practical laboratories 421 Catalan/Spanish second semester morning-mixed
(PLAB) Practical laboratories 422 Catalan/Spanish second semester morning-mixed
(PLAB) Practical laboratories 423 Catalan/Spanish second semester morning-mixed
(PLAB) Practical laboratories 424 Catalan/Spanish second semester morning-mixed