General Information

·         History

·         Fundamentals / general info

·         DNA Cloning / principles of Recombinant DNA technology

·         Cloning using E.coli

·         Restriction modification of DNA

Restriction Modification

 ·         Basic concepts / definitions

·         General example

·         Specific examples

·         Restriction endonucleases; classes, properties

·         Enzymes used in recombinant DNA technology

Cloning Vectors

·         Plasmids

·         Bacteriophage

·         Cosmids

 Characterizing DNA

·         Density gradient centrifugation

·         Electron microscope

·         Electrophoresis

·         Non-enzymatic cleavage

·         Enzymatic cleavage

·         Restriction Mapping (general rules / practical considerations / methodology) 

DNA Sequencing

 ·         Gilbert-Maxam method

·         Sanger-Dideoxy Method

·         DNA library

·         Audioradiography / hybridization

·         Forensic analysis

·         Restriction Fragment Length Polymorphisms  (RFLP)

·         DNA fingerprinting

 PCR - Polymerase Chain Reaction

 ·         General information

·         History of PCR development

·         Mechanics of PCR (denaturation, primer annealing, primer extension)

·         Primers

·         Annealing details

·         PCR specificity (temperature, concentration, time, mismatches)

·         Taq Polymerase

 Automated PCR

 ·         Thermal cycler

·         Automated PCR preparation

·         Sensitivity and unwanted contaminants

 Example PCR Methodology

·         Sample components / concentrations

·         Typical cycle conditions

·         Fast / Turbo PCR

·         Clean-up and amplicon storage

·         Direct restriction enzyme digestion

Inverse PCR

 ·         Concept

·         Process flow

Anchor PCR

 ·         Concept

·         Process flow

Sequencing DNA Directly after PCR 

·         Assymetric Primer ratios

·         Biotinylated primers

 PCR Applications 

·         Mutations, genetic disorders. Oncogenes, pathogens, forensics, fastidious cultures

·         Advantages / disadvantages of PCR

 Molecular Diagnosis of Infectious Diseases

·         General information

·         Microbial phenotype vs. Nucleic acid techniques

Traditional Microbial Testing

 ·         Biotyping

·         Antibiotic reseistance / Resistograms

·         Protein analysis

·         Phage analysis

·         Chromatography

Nucleic Acid Typing Systems

·         Plasmid analysis

·         Restriction RFLP analysis

·         Ribotyping

·         RAPD (random amplified polymorphic DNA analysis)

 Nucleic Acid Analysis without Amplification 

·         Nucleic acid probes

·         Branched DNA signal amplification

 PCR and Diagnostics

·         Reverse transcriptase PCR

·         Nested PCR

·         Multiplex PCR

·         Broad Range PCR

 Other Nucleic Acid AmplificationTechniques 

·         TAS (transcription-based amplification system)

·         LCR (ligase chain reaction)

·         SDA (strand displacement amplication)

·         QB Replicase system

 Analysis of Amplification Products 

·         Conventional nucleic acid amplification

·         HPA (hybridization protection assay)

·         DEIA (DNA enzyme immunoassay)

·         Automated DNA sequencing technology

·         SSCP (single strained conformational polymorphisms)

·         RFLP (restriction fragment length polymorphisms

·         Summary of nucleic acid diagnostics