Molecular Biology Summer Workshops

Course Lecture Topics and Laboratory Experiments

 

Lecture Topics

  • 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
  • Next Generation DNA sequencing and RNAseq
  • Cloning in Eukaryotic Cells
  • Cloning in Cosmids and YAC Vectors
  • Immunoscreening Libraries
  • Gene Expression/Protein Production in Heterologous Hosts
  • DNA Fingerprint Analysis
  • Genomics and Bioinformatics

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

BRIEF SUMMARY OF THE FIVE MOLECULAR BIOLOGY PROJECTS
  Experiment #1: Genomic Cloning and Protein Expression
  • cDNA cloning of the mouse GAPDH gene in a plasmid expression vector (pMAL).
  • First and second strand cDNA synthesis and PCR to synthesize the GAPDH gene.
  • Ligation of the GAPDH cDNA into the plasmid expression vector pMAL.
  • Transformation of E. coli, selection of recombinant clones and DNA sequencing.
  • Expression and purification of the GAPDH gene fusion in E. coli.
  • Measure protein concentration and run on protein gels.
  • Western blot to specifically detect the GAPDH fusion protein.
Experiment #2: Genome Analysis
  • Isolate and purify mouse genomic DNA from liver tissue.
  • Amplify the transthyretin (Ttr) gene from total mouse DNA using the polymerase chain reaction (PCR).
  • Use biotin-labeled oligonucleotide probes for hybridization to the Southern blot.
  • Southern blot analysis of the Ttr and Rvt genes from mouse DNA.
  • Analysis of the methylation state of the Rvt genes in mouse genomic DNA.
Experiment #3: Gene Expression Analysis & RNA seq
  • Preparation of total RNA from mouse liver tissue
  • Amplification of transthyretin (Ttr) mRNA by reverse transcriptase PCR (RT-PCR)
  • Quantitative RT-PCR using real-time analysis (TaqMan system)

Experiment #4: Next Generation Sequencing(NGS): RNAseq to Study Gene Expression in Mouse Liver

  • Library construction for RNAseq
  • RNAseq bioinformatics

Experiment #5: RNA interference in C. elegans

  • RNA interference (RNAi) in C. elegans

Experiment #6: Human DNA/Genetic Analysis

  • Isolation of your own DNA from cheek cells from your mouth.
  • PCR amplification of your own DNA for DNA fingerprint analysis.
  • PCR amplification of your taster gene: compare phenotype and genotype.
  • DNA fingerprint analysis.
 
 

 

 

   
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  This  page is maintained by Susan Haynes at the Ford Hall, Smith College, Northampton, MA 01063.  Last update: January 25, 2016.