Molecular Biology Summer Course Lecture
Topics (Partial List)
Enzymes and Ligase Enzymes for Cloning DNA
- DNA Manipulations: Linkers and Adaptors
- Cloning Vectors: Plasmids and Bacteriophage
- Bacteriophage Lambda
- Genetic Selection Techniques and Cloning
- Genomic Library Construction (Including
- cDNA Library Construction (Including
- 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
- DNA/DNA and DNA/RNA Hybridization
- Chain Termination (Dideoxy) DNA Sequencing
- Thermal Cycle Sequencing
- Automated and Non-radioactive DNA
- Computer Analysis of DNA Sequences
- Pulse Field Gel Electrophoresis
- Gene Expression and Regulation in
- Cloning in Eukaryotic Cells/Transgenic
- Cloning in Cosmids and YAC Vectors
- Immunoscreening Libraries/Immunological
- Gene Expression/Protein Production in
Note: Some of these topics will be combined in a single
lecture, while others will take several lectures.
Molecular Biology Summer Course 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 to a plasmid cloning vector.
- Isolation of recombinant plasmid DNA
- Dideoxy DNA sequencing (thermal cycle and
non-radioactive) of the cloned mouse gene.
- Computer analysis of the DNA sequence data.
Experiment #2: Genome Analysis
- Isolate and purify mouse genomic DNA from
liver and kidney tissues.
- Amplify the prealbumin gene from total
mouse DNA using the polymerase chain reaction (PCR).
- Non-radioactive 3' end labeling of a
prealbumin oligonucleotide probe and random prime labeling of the mouse
reverse transcriptase gene.
- Southern blot analysis of the mouse
prealbumin gene and the reverse transcriptase gene.
Experiment #3: Gene Expression Analysis
- Preparation of mRNA from mouse liver total
- Amplification of prealbumin mRNA by reverse
- RT-PCR analysis of gene expression.
- Quantitative RT-PCR analysis of gene
- RNase protection assays.
Experiment #4: cDNA Cloning and PCR Library Analysis
- Construction of a unidirectional cDNA
library from mouse liver mRNA in a lambda bacteriophage expression
- Screening the cDNA library by PCR and
- Analysis of cDNA insert size and the
prealbumin gene by PCR.
was created and maintained by Dr. Wen Li
at the Clark Science Center, Smith College, Northampton, MA
01063. Last update: October 30, 2003.