Molecular Biology

(Three workshops during the year 2008)

Organized by: Dr. Eman El-Abd

NB. Enrollment limited to 15 participants

Level I: Basic Molecular Biology

This workshop aims to introduce Scientists and Clinicians to molecular isolation and characterization of DNA, RNA, and protein. This program will be a requisite for the forth-coming program (basic molecular biology level II) which will be mainly about PCR and Western analysis. 

Day 1:

09:00 ・10:00               DNA structure and function

10:00 ・11:00               DNA extraction

11:00 ・11:30               Break

11:30 ・02:00               Practical session (DNA extraction from whole blood)

Day 2:

09:00 ・10:00               RNA structure and function

10:00 ・11:00               RNA extraction

11:00 ・11:30               Break

11:30 ・02:00               Practical session (RNA extraction from tissue)

Day 3:

09:00 ・11:00               Protein extraction and characterization

11:00 ・11:30               Break

11:30 ・02:00               Practical session I (protein extraction)

Day 4:

09:00 ・11:00               Practical session II (SDS-PAGE)

11:00 ・11:30               Break

11:30 ・ 02:00               Practical session III (Staining, distaining, visualization)

Level II: PCR Techniques and Western Blot Analysis

Day 1:

09:00 ・10:00               Introduction to PCR Theory

10:00 ・10:30               Break

10:30 ・11:30               Nested PCR and Multiplex PCR

11:30 ・12:00               Practical session

Day 2:

09:00 ・10:00               RT-PCR

10:00 ・10:30               Break

10:30 ・11:30               Mutation detection using PCR (RFLP, SSCP, AS, LCR)

11:30 ・12:00               Practical session

Day 3:

09:00 ・10:00               Other types of PCR (Long PCR, Asymmetric PCR, Inverse PCR....etc)

10:00 ・10:30               Break

10:30 ・11:30               Real time PCR

11:30 ・12:00               Detection of TB by PCR

Day 4:

09:00 ・10:00               Principles of Western blot analysis

10:00 ・10:30               Break

10:30 ・12:30               Practical session (SDS-PAGE, blotting)

Day 5:

09:00 ・10:00               Blocking of the non specific binding

10:00 ・10:30               Break

10:30 ・11:30               Hybridization with the first antibody

Day 6:

09:00 ・10:00               Hybridization with the second antibody

10:00 ・10:30               Break

10:30 ・11:00               Detection of signal 

Level III: Subcloning

This workshop teaches advanced methods of gene manipulation, and characterization. An assortment of the following techniques will be covered: subcloning DNA fragments into plasmid vectors and characterization of DNA clones restriction digestion analysis. Prerequisite: Basic knowledge of nucleic acids (level I and II or equivalents) is advantageous.

Day 1:

                                    Lecture session

09:00 - 11:00               Sub-cloning: importance and strategies

11:00 - 11:30               Break

12:00 - 02:00               Practical session

                                    Preparation of cDNA (insert)

Day 2:

                                    Lecture session

09:00 - 11:00               Vectors and Enzymes used in molecular sub-cloning

11:00 - 12:00               Bacterial strains, media and antibiotics

12:00 - 12:30               Break

01:00 - 02:00               Practical session

Preparations of media and competent E. coli

Day 3:

                                    Lecture session

09:00 - 10:00               Dephosphorylation and strategies for ligation

10:00 - 11:00               Transformation and screening strategies

11:00 - 11:30               Break

11:30 - 02:00               Practical session

Ligation and transformation

Day 4:

                                    Practical session

09:00 - 11:00               Identification of colonies with recombinant plasmids

                                    Rapid small-scale preparation of plasmid DNA

11:00 - 11:30               Break

11:30 - 02:00               Restriction digestion of plasmid DNA

Gel electrophoresis

Setting an overnight bacterial culture

Day 5:

                                    Practical session

09:00 - 11:00               Harvesting and lysis of bacteria

                                    Large-scale preparation of plasmid DNA

11:00 - 11:30               Break

11:30 - 02:00               Purification of plasmid DNA