DNA-PROFILING AND FORENSIS GENETICS
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- Versione italiana
- Academic year
- 2021/2022
- Teacher
- ELISABETTA MAMOLINI
- Credits
- 6
- Curriculum
- BIODIVERSITA' ED EVOLUZIONE
- Didactic period
- Secondo Semestre
- SSD
- BIO/18
Training objectives
- Genetic-forensic field, starting from a specific biological trace, the ideal perspective is to obtain a complete DNA profile and high quality. The genetic traceability, is also taking an important contribution to the control of quality and safety of agricultural raw materials and processed products, as it allows to evaluate the authenticity of a product. The analysis of short nucleotide sequences of mitochondrial is undoubtedly become a widely used method in the identification of plant and animal species, in forensics (DNA barcoding) and is developing in the diagnosis of human diseases. Mitochondrial genes are used as barcode sequences since the mtDNA has a high number of copies of mtDNA present in the cells facilitates the recovery of degraded tissues and also by PCR amplification. The DNA Barcoding has proved particularly useful in food safety, enabling the identification of species even in processed products in which the morphological characteristics of the organisms are no longer recognizable. The main objective of the Course is to provide students with the knowledge to study and understand the practical applications of systems based on DNA Profiling and DNA Barcoding, for the understanding of the relationship between the forensic genetics and its latest applications in the field of biomolecular and evolutionary.
KNOWLEDGE
the student:
He knows the genetic terminology correctly;
He knows the biological basis of hereditary processes
knows the basics of basic cytogenetics (mitosis and meiosis)
He knows human forensic genetics, animal and plant
He knows the most used computer sites in forensic genetics
He knows how to search for references in international databases
He knows the techniques to extract DNA from different biological matrices
CAPACITY 'TO APPLY KNOWLEDGE AND UNDERSTANDING
the student:
You know how to properly use the genetic terminology;
You know how to evaluate the possible impact of forensic genetics in its reality,
sa look in the bibliography forensic sites suitable
He knows how to design an experiment
He knows the techniques to extract DNA from human traces, plant and animal. Prerequisites
- There is no rule in the prerequisites, however, the student must have the following knowledge of genetics and biology: DNA, replication, inheritance (mitosis and meiosis), allele and genotype frequencies
Course programme
- Forensic Genetics (14 hours):
STRUCTURE AND FUNCTIONS OF DNA
- Basics of cytology
- Structure of DNA, genes and non-coding DNA
-Mutations and polymorphisms of DNA in forensic genetics (VNTR, STR, SNP), DNA fingerprinting and DNA profiling; CODIS
-polymorphisms of the Y chromosome and mitochondrial DNA (mtDNA)
- database in genetics forensics (nucleotide, and haplotype DNA profiling)
MOLECULAR GENETICS: METHODS OF ANALYSIS OF GENETIC VARIABILITY AT THE DNA LEVEL
- biological traces (blood, saliva, etc.) techniques and technologies of the inspection
Tests for personal identification (classical hemogenetics, generic species diagnosis, ordinary DNA sources)
Techniques for the analysis of polymorphisms
- PCR end-point
- DNA sequencing
-Real Time PCR (qPCR)
Analysis of the results:
- Allelic assignment and determination of the genotype and software used
- Interpretation of electropherograms: Drop-in; Drop-out; Stutter bands
Statistics applied to genetic-forensic investigations:
Frequency distributions; Estimation of confidence intervals; Significance test; Estimation of allele and genotype frequencies; Estimation of allele frequencies to codominant loci (VNTR or STR); Estimation of genotypic frequencies to multilocus systems
Identification; Probability of exclusion; Probability of match; Probability of identity;
Paternity test
The role of Ethics in forensic investigations
Analysis of the results:
- Allelic assignment and determination of genotype and software
-interpretation of electropherograms: Drop-in; Drop-out; Stutter bands
Statistics applied to the genetic-forensic investigations:
Frequency distributions; confidence intervals; Significance tests; allele and genotype frequencies; Allele frequencies at codominant loci (VNTR or STR); estimate genotypic frequencies in multi-locus systems
Identification; Probability of exclusion; Likelihood of match; Probability of identity; paternity test
The role of ethics in forensic investigations
(10 hours) NGS; DNA Barcoding (Standard barcode sequences: the Cytochrome oxidase I (COI), the Cytochrome b and 16S),
Applications in:
1) epidemiology and public health;
2) Consumer protection;
3) Identification of the species present in fresh or processed foods, tracking during production, processing, packaging and marketing;
4) Customs Regulations;
5) Environmental monitoring.
Barcode of Life Data Systems (BOLD) site that organizes all the sequences of living beings (www.boldsystems.org).
Laboratory (24 hours-2 CFU)
Consultation of DNA databases: genetic databases for forensic use (nucleotides, haplotypes, genetic profiles, CODIS and Y-chromosome and mitochondrial DNA polymorphisms);
Cytological analysis (microscope) identification of the human blood group (ABO and Rh);
Forensic human genetic investigations:
Extraction of DNA from human buccal cells (plastic bottle residue); from cigarette butts;
DNA extraction from blood
Analytical investigations of foods:
DNA extraction from wheat (liquid nitrogen-kit);
DNA extraction from animal meat (phenol-chloroform);
DNA extraction from fish
DNA quantification (NanoDrop)
Qualitative PCR or End Point; PCR multiplex
Preparation of agarose gel;
Visualization of the results and determination of the genotype
Quantitative PCR or Real Time PCR (qPCR) for the identification and quantification of species and / or contamination present in food. Didactic methods
- The course is structured in theoretical lessons for a total of 24 hours of teaching (3+1 credits). The lessons will take place in presence and weekly by registering them with the Classroom platform, using slides on power-points; for the best understanding of the topics and to answer your possible questions, direct interaction with Google meet will be used. About 10 hours of the course will be assigned to a colleague to deal with the following topics: NGS; Barcoding DNA.
24 hours (2 credits), only if possible, will be dedicated to the exercises that will take place in the laboratory in the presence of the students. Learning assessment procedures
- The exam consists of written or a telematic oral exam test aimed at assessing the student's theoretical abilities in relation to the biological-biomolecular approach of Forensic Genetic Sciences.
The outcome of the exam will be out of thirty: the test is deemed passed with a mark of at least 18/30 of the test.
The exam is oral (Google meet) and consists of 3-4 questions, the answers of which will be judged on the basis of both the content and the appropriate scientific language. The maximum score for each is 9 points. The duration of the whole test will be approximately 15 minutes minutes. The exam is not passed if the overall assessment is less than 18/30.
The test allows to ascertain the achievement of the objectives in terms of knowledge and applied comprehension skills. The exam also aims to understand oral communication skills in relation to the use of specific language. Reference texts
- Introduction to Forensic Genetics: personal identification and paternity investigations. Tagliabracci- Springer.
The DNA test and the biological proof of paternity. Marina Dobosz- PICCIN
Genetic-Forensic investigations of paternity and personal identification (2) by Tagliabracci, Domenici, Pascali, Pesaresi. PICCIN
They will also be made available for students slides of lectures.
