BIOC 2203L Syllabus

Subject Code

BIOC

Course Number

2203L

Course Title

Recombinant DNA Methods Lab

Prerequisites

BTEC 2192 with a grade of C or higher; BTEC 2192L with a grade of C or higher

Corequisites

BIOC 2203

Terms Offered

Credit Hours

Course Description

This recombinant DNA laboratory course provides students with fundamental molecular techniques involved in genetic engineering. Intensive bench training includes large scale plasmid isolation, restriction analysis, ligations, generation of recombinant DNA, preparation of a genomic library, southern blot analysis, and purification of a restriction enzyme. Furthermore, students will develop and perform PCR protocols as part of a research project analyzing a selected class of genetically modified organisms. The research project must be accompanied by authoring a formal research report to be presented in class.

Course Outcomes

To Prepare A Recombinant Plasmid Conferring Double Antibiotic Resistance In E. Coli

To prepare large scale plasmid DNAs.
To prepare specific restriction digests and ligation products.
To perform a transformation of an appropriate host strain with the recombinant ligation mixture.
To explain the transformation results yielding a specific recombinant plasmid structure using proper selection.

To Generate A Phage Library Using Blue-White Cloning

To prepare special media containing a chromogenic substrate.
To prepare restriction digests and ligation products using appropriate vector DNA and specific insertion fragments.
To perform a blue-white cloning procedure.
To examine a phage library construct.

To Purify and To Assay A Restriction Enzyme

To demonstrate the ability to pack and to equilibrate DEAE-cellulose columns.
To perform the purification of EcoRI.
To examine purified EcoRI using assays verifying and documenting its specific restriction activity.

To Perform A Southern Blot Analysis

To demonstrate the ability to transfer DNA fragments from a gel onto a membrane.
To describe the design of sequence-specific DNA probes for detection.
To describe non-isotopic detection systems using specific biotinylated probes.
To point out DNA-DNA hybridization products using specific protocols for color development.

To Conduct and To Analyze PCR Reactions Based On Standard Protocols

To demonstrate the ability to perform PCR reactions using standard protocols such as for genetic typing, reverse transcriptase and water quality testing.
To examine reverse transcriptase PCR results.
To examine PCR-based water quality testing results for coliforms.
To examine PCR-based Alu-human DNA typing results.

To Develop, Conduct and To Complete An Independent PCR Project

To describe specific primer design required to amplify loci detecting genetically modified maize and/or soy.
To develop specific PCR protocols allowing the detection of genetically modified plant DNAs.
To demonstrate the ability to perform PCR reactions based on developed protocols.
To examine PCR results testing maize and/or soy genomes for the presence of genetically modified DNAs.
To produce a unique laboratory report documenting PCR research results.