Lecture Topics (Two-Week Courses)

• RNA interference, siRNA and microarray analysis
• Restriction Enzymes and Ligase Enzymes for Cloning DNA
• DNA Manipulations: Linkers and Adaptors
• Cloning Vectors: Plasmids and Bacteriophage
• Bacteriophage Lambda
• Genetic Selection Techniques and Cloning Strategies
• Genomic Library Construction
• cDNA Library Construction
• Polymerase Chain Reaction (PCR)
• Reverse Transcriptase PCR (RT-PCR)
• Quantitative Real-Time PCR
• Gel Electrophoresis
• DNA and RNA Isolation and Purification
• Southern Blot Analysis
• Analysis of Gene Expression
• Northern Blot Analysis
• In situ Hybridization Analysis
• DNA Labeling Methods (Radioactive and Non-radioactive)
• DNA/DNA and DNA/RNA Hybridization
• Chain Termination (Dideoxy) DNA Sequencing
• Thermal Cycle Sequencing
• Automated and Non-radioactive DNA Sequencing
• Computer Analysis of DNA Sequences
• Pulse Field Gel Electrophoresis
• Gene Expression and Regulation in Mammalian Systems
• Cloning in Eukaryotic Cells
• Cloning in Cosmids and YAC Vectors
• Immunoscreening Libraries
• Gene Expression/Protein Production in Heterologous Hosts
• DNA Fingerprint Analysis
• Bioinformatics

Note: Some of these topics will be combined in a single lecture, while others will take several lectures.

• Construction of a size-selected mouse genomic library in the bacteriophage vector Lambda ZAP Express
• Non-radioactive labeling of a mouse repeat DNA probe using PCR
• Selection of a specific mouse repeated gene (reverse transcriptase, Rvt repeat) from this library with the non-radioactive DNA probe.
• PCR subcloning of the Rvt repeat gene from the lambda cloning vector into a plasmid cloning vector
• Isolation of recombinant plasmid DNA (plasmid prep) from colonies
• Dideoxy DNA sequencing (thermal cycle and non-radioactive) of the cloned Rvt gene.
• Computer analysis of the DNA sequence data (bioinformatics).

• Isolate and purify mouse genomic DNA from liver tissue.
• Amplify the transthyretin (Ttr) gene from total mouse DNA using the polymerase chain reaction (PCR).
• Quantification of genomic DNA using spectrophotometry and gel electrophoresis methods

• Preparation of RNA from mouse liver tissue
• Amplification of transthyretin (Ttr) mRNA by reverse transcriptase PCR (RT-PCR)
• RT-PCR analysis of gene expression
• Quantitative RT-PCR using real-time analysis of gene expression (TaqMan system)
• RNA interference (RNAi) in C. elegans
• Microarray analysis and bioinformatics

• Expressing a gene fusion to produce a reporter construct
• Purify the expressed protein

• Isolation of your own DNA from cheek cells. (Just like on CSI!)
• Multiplex PCR amplification and fingerprint analysis of your own DNA for fingerprint analysis
• Mitochondrial DNA analysis and molecular evolution of the mitochondrial genome
• PCR amplification of your taster gene: compare phenotype and genotype
• DNA fingerprint analysis and DNA sequencing of your mitochondrial on the ABI 310 Gene Analyzer

• Analyses of gene silencing by observing phenotypes in C. elegans.
• RNAi and bioinformatics.