Lecture Topics (Partial List for 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 (Including Size Selection)
• cDNA Library Construction (Including Subtraction Libraries)
• Polymerase Chain Reaction (PCR)
• Reverse Transcriptase PCR (RT-PCR)
• Quantitative 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/Transgenic Animals
• Cloning in Cosmids and YAC Vectors
• Immunoscreening Libraries/Immunological Applications
• 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 DNA probe using PCR.
• Selection of a specific mouse gene (reverse transcriptase) from this library with the non-radioactive DNA probe.
• PCR subcloning of the mouse gene from the lambda cloning vector into a plasmid cloning vector.
• Isolation of recombinant plasmid DNA (plasmid prep).
• Dideoxy DNA sequencing (thermal cycle and non-radioactive) of the cloned mouse gene.
• Computer analysis of the DNA sequence data (bioinformatics).

• Isolate and purify mouse genomic DNA from liver tissue.
• Amplify the transthyretin gene from total mouse DNA using the polymerase chain reaction (PCR).
• Non-radioactive 3' end labeling of a transthyretin oligonucleotide probe.
• Southern blot analysis of the mouse transthyretin gene and the reverse transcriptase gene.

• Preparation of RNA from mouse liver.
• Amplification of transthyretin mRNA by reverse transcriptase PCR.
• RT-PCR analysis of gene expression.
• Quantitative RT-PCR analysis of gene expression.
• Microarray analysis.
• Sequence analysis of Ttr gene and bioinformatics.

• Expressing a gene fusion to produce a reporter construct.
• Purify the expressed protein.
• Western Blot transfer.
• Monoclonal antibody detection of the recombinant protein.

• Isolation of your own DNA from cheek cells. (Just like on CSI!)
• Multiplex PCR amplification and fingerprint analysis of your own DNA.
• Mitochondrial DNA analysis and molecular evolution of the mitochondrial genome.

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

• The beginner one week course (Session 3) will complete experiments #1, #2 and #3 along with coordinating lectures.

• The advanced one week course (Session 4) will complete experiments #4, #5 and #6 with appropriate coordinating lectures.