FEBS Course: Recombinant DNA Technology

septembrie 2, 2001 - septembrie 7, 2001

  • FEBS Course: Recombinant DNA Technology

    • septembrie 2, 2001 - septembrie 7, 2001
    • Bucharest

    FEBS Advanced Theoretical and Practical Course:

    RECOMBINANT DNA TECHNOLOGY

    September 2-7, 2001

    Bucharest, Romania

    Accomodation & Registration

    The participants will be accommodated in double rooms at the Academica Hotel, which is situated in the Center of Bucharest, near the Law Faculty and the Opera House. Registration will take place at the Hotel, in the evening of 1st September, between 5 and 10 pm.

    How to reach the Hotel:

    From the Otopeni International Airport

    The shuttle bus # 783 links the airport with the Center of Bucharest. A card for 2 trips costs 20,000 lei and you can buy it from the driver. You have to get off the bus at the “Universitatii” Station (University Station) and change with one of the trolleys # 69 or 90 or bus # 336. A ticket for one trip with the trolley or the bus costs 5,000 lei and you can buy it from the yellow boxes marked RATB in the stations. You have to get off the trolley or bus at “Facultatea de Drept” Station (Law Faculty Station). The Hotel is right in front of the station, at the end of a short alley.

    You can go directly from the Airport to the Hotel by taxi. We strongly advise you to take a yellow taxi belonging to one of the taxi companies: “Cobalcescu”, “Alfa” or “Meridian”. The trip will cost you about 250,000 lei.

    From the main railway station “Gara de Nord” ( Northern Railway Station)

    You have to take either the bus # 123 or the subway. You can take the bus from the front of the Railway Station. A ticket for one trip with the bus costs 5,000 lei and you can buy it from the yellow boxes marked RATB in the stations. You have to get off at the “Medicina Veterinara” Station (Veterinary Medicine Faculty Station) and cross by foot the Opera Bridge across the Dambovita River. At the other end of the bridge you will see the bus station “Facultatea de Drept” (Law Faculty Station). The Hotel is right in front of the station, at the end of a short alley.

    The subway station (Metro – sign “M”) can be easily reached within the Railway Station, by means of descending stairs. A card for 2 trips costs 10,000 lei and you can buy it from the subway station. You have to take the train in direction “Republica” and get off at the “Eroilor” Station (Station of Heroes). You have to leave the station through the “Piata Operei” (Opera Square) exit and cross by foot the Opera Bridge across the Dambovita River and pass the Opera Building. You will see the bus station “Facultatea de Drept” (Law Faculty Station). The Hotel is right in front of the station, at the end of a short alley.

    You can go directly from the Railway Station to the Hotel by taxi. We strongly advise you to take a yellow taxi belonging to one of the taxi companies: “Cobalcescu”, “Alfa” or “Meridian”. The trip will cost you about 75,000 lei.

    Location of the course:

    Institute of Biochemistry of the Romanian Academy, Bucharest, Romania.

    The Institute has the necessary laboratory space, equipment and special halls for seminars and lectures. A nice garden surrounding the institute building could be used for walking and social events, eventually.

    Participants

    Invited lecturers:

    Dr. Hans Lenstra, Institute of Infectious Diseases and Immunology, Utrecht University, The Netherlands:

    Lecture title: “A Decade of Revolution in Molecular Genetics, 1990-2000

    Professor Ami Aronheim, Dept. Molecular Genetics and the Rappaport Family Institute, Faculty of Medicine, Haifa, Israel

    Lecture title: “Protein Recruitment Systems for the Analysis of Protein-Protein Interactions

    Dr. Panagiotis Deloukas, Dept. of Human Genetics, The Sanger Centre, Wellcome Trust Genome Campus, Cambridge, UK

    Lecture title: “Towards a Finished Reference Sequence of the Human Genome

    Professor Ralph Dornburg, The Dorrance H. Hamilton Laboratories, Thomas Jefferson University, Philadelphia, USA

    Lecture title: “Retroviral Vectors in Human Gene Therapy

    Tutors:

    Dr. Gabriela Anton, Institute of Virology, Bucharest

    Dr. Norica Nichita, Institute of Biochemistry of the Romanian Academy, Bucharest

    Dr. Adrian Onu, Cantacuzino Institute, Bucharest

    Dr. Dan Otelea, Cantacuzino Institute, Bucharest

    Prof. Octavian Popescu, Babes-Bolyai University, Cluj-Napoca

    Dr. Rodica Repanovici, Institute of Virology, Bucharest

    Dr. Stefan Szedlacsek, Institute of Biochemistry of the Romanian Academy, Bucharest

    Students:

    20 participants have been finally accepted, of which 13 from abroad are supported by Youth TravelFunds (YTF).

    5 Romanian students are partially supported by The Romanian Society of Biochemistry and Molecular Biology.

    Most of the participants are Ph.D. students.

    Organizing committee:

    • Dr. J. A. Lenstra, Institute of Infectious Diseases and Immunology, Utrecht University, The Netherlands
    • Dr. Stefan Szedlacsek, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    • Dr. Stefana Petrescu, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    • Dr. Gabriela Negroiu, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    • Dr. Anca Roseanu, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    • Dr. Rodica Repanovici, Institute of Virology, Bucharest
    • Dr. Gabriela Anton, Institute of Virology, Bucharest
    • Ph.D. Student Alexandra Hillebrand, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    • Ph.D. Student Vlad Pena, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania
    • B.Sc. Catalin Lazar, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania

    Aim of the course:

    The aim of this course is to help the young biochemists to acquire the basic skills in manipulating DNA, building recombinant DNA and expressing recombinant proteins in the prokaryotic system. This course is intended to offer to participants an introduction to the state-of-the-art techniques in molecular biology.

    Final Scientific Programme:

    Day 1, Sunday September 2

    09.00

    Welcome

    Introduction to the course

    09.30

    Tutorials

    Techniques of DNA and RNA isolationDr. Gabriela Anton

    DNA electrophoresisDr. Dan Otelea

    Polymerase Chain Reaction (PCR)Dr. Rodica Repanovici

    Reverse transcription and cDNA librariesDr. Gabriela Anton

    13.00

    Lunch

    14.30

    Experiments

    -Purification of total RNA from B16 cells

    -Obtaining cDNA from total RNA using reverse transcription

    19.00

    Invited lecturer: 

    Professor Hans Lenstra:

    A decade of revolution in molecular genetics, 1990-2000

    Day 2, Monday September 3

    09.00

    Tutorials

    Southern and Northern blottingDr. Rodica Repanovici

    09.30

    Experiments

    -Southern blot

    -RT-PCR (one step)

    13.00

    Lunch

    14.30

    Tutorials

    RestrictionDr. Dan Otelea

    15.30

    Experiments

    -Southern blot (cont.)

    -Gel extraction of RT-PCR product

    -Hybridization (over night) of Southern blot membrane with DIG-labeled probe

    19.00

    Invited lecturer: 

    Prof. Ami Aronheim

    Protein recruitment systems for the analysis of protein-protein interactions

    Day 3, Tuesday September 4

    09.00

    Tutorials

    DNA ligationDr. Norica Nichita

    DNA transformation” Dr. Norica Nichita

    10.00

    Experiments

    -Plasmid purification 

    -DNA Gel electrophoresis for visualization of RT-PCR product and purified plasmid

    -Digestion with restriction enzymes of both RT-PCR product and cloning vector 

    13.00

    Lunch

    14.30

    Tutorials

    VectorsDr. Adrian Onu

    15.30

    Experiments

    -Gel extraction of digested DNA

    -DNA Gel electrophoresis

    -Ligation (over night 16ºC)

    19.00

    Invited lecturer: 

    Professor Ralph Dornburg 

    Retroviral vectors in Human Gene Therapy

    Day 4, Wednesday September 5

    09.00

    Tutorials

    Site-directed mutagenesisDr. Adrian Onu

    10.00

    Experiments

    -Transformation by electroporation of ligation product

    -Plating out of transformed E.coli

    11.30

    Tutorials

    DNA sequencingProfessor Octavian Popescu

    13.00

    Lunch

    16.00

    Social Event

    Day 5, Thursday September 6

    09.00

    Tutorials

    Exercises on DNA chemistryProfessor Octavian Popescu

    10.00

    Experiments

    -Counting of colony

    -Screening by PCR

    13.00

    Lunch

    14.30

    Tutorials

    Bioinformatics IDr. Stefan Szedlacsek

    15.30

    Experiments

    -Gel electrophoresis for visualization of positive clones

    -Overnight culture of positive clones

    19.00

    Invited lecturer: 

    Prof. Panagiotis Deloukas

    Towards a finished reference sequence of the human genome

    Day 6, Friday September 7

    09.00

    Experiments

    -Purification of recombinant plasmid

    -Visualization of recombinant plasmid in agarose gel

    10.30

    Discussions with tutors

    13.00

    Lunch

    14.30

    Tutorials

    Bioinformatics IIDr. Stefan Szedlacsek

    18.30

    Final evaluation, closure

    Useful tips

    The weatherin Bucharest in September is generally sunny, temperature ranging 15-25 degrees Celsius. However do not forget to take your umbrella, it might be useful.

    If you are a student, don’t forget to take your student card(preferably an international student card) with you. You will find that in Romania, students pay smaller prices for some services (e.g. to visit museums).

    National currencyof Romania is LEI. The exchange rate is approximately:

    1 USD = 30,000 lei

    1 Euro = 27,500 lei

    1 DEM = 14,000 lei

    There are many exchange officesin Bucharest. On the Airport and in the Railway Station you can also find them, but we advice you to avoid exchanging your money in these two places because the exchange rates are unfavorable as compared to other exchange offices in the city.

    Romanians are generally considered by foreigners as friendly and ready to help people coming from abroad. So don’t hesitate to ask people’s help whenever you need assistance. Especially the young people can speak English.

    Contact

    Once in Bucharest, don’t hesitate to contact us if you need any help.

    Institute of Biochemistry:

    Splaiul Independentei, 296

    Phone: 223 90 69

    223 90 68

    Hotel Academica:

    Bd. Regina Elisabeta 34-44

    Phone: 310.42.69

    Mobile phones:

    Vlad Pena 

    Lucian Moldovan

    094 22 31 89

    093 98 26 48

    Last update: August, 21, 2001

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