Molecular Biology Summer Workshops

Course Lecture Topics and Laboratory Experiments

 

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.

Laboratory Experiments (Partial List)
  Experiment #1: Genomic Cloning and DNA Sequencing
  • 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).
Experiment #2: Genome Analysis
  • 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.
Experiment #3: Gene Expression Analysis
  • 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.
Experiment #4: Protein Expression, Purification and Western Blot Analysis 
  • Expressing a gene fusion to produce a reporter construct.
  • Purify the expressed protein.
  • Western Blot transfer.
  • Monoclonal antibody detection of the recombinant protein.
Experiment #5: DNA Fingerprint Analysis
  • 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.
Experiment #6: RNA Interference
  • Analyses of gene silencing by observing phenotypes in C. elegans.
  • RNAi and bioinformatics.
One Week Courses
  • 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.
   
  BACK HOME
   
  This  page is maintained by Susan Haynes at the Clark Science Center, Smith College, Northampton, MA 01063.  Last update: April 14, 2008.