Techniques in Molecular Diagnostics:

2 credits - Spring Semester
Course Director - Joe Nickels, Ph.D.

This course is designed to allow students to master techniques routinely used in molecular diagnostics.  Students will develop and apply these techniques in a laboratory-based setting. Methods include DNA and RNA isolation and quantification; protein expression, purification and analysis; molecular cloning methods; diagnostic methods used for genetic testing; culture methods for growth of bacteria, yeast and viruses; microscopic methods for diagnostic testing, etc. Students will select two techniques during the course to perform and master.  Students will be required to write a short (4-5 page) NIH-type introduction on each method.  In addition, students will give an oral presentation on one of the techniques mastered. Upon completion of this course students will have acquired a basic mastery of a subset of methods routinely used in the molecular diagnoses of disease.

Course Outline:

Summer Semester
Week	Topic
1	Introduction
1 - 7	Laboratory – Technique #1
8	NIH type introduction on Technique #1	(4-5 pgs)
8 - 14	Laboratory – Techniques #2
15	Student presentations
15	NIH type introduction on Technique #2	(4-5 pgs)

Grading:
Laboratory attendance and attendance at final oral presentations of all students is mandatory. Each NIH type introduction and individual student evaluations will be graded by the faculty member heading the laboratory the student chooses for techniques 1 and 2.  Grades will be based upon completion of all projects as follows:
Written assignments (2)	40% total
Evaluation by laboratory mentor (2)	40% total
Oral Presentation	20%
Below is a list of the techniques that are available to students. The students will choose two techniques to master.  Each "technique rotation" will be seven weeks. In addition to submitting a 4-5 page NIH-style introduction on each technique, students will choose one of the techniques for their oral presentation.
John Blaho, Ph.D Growth, storage, and quantitation of human herpes simplex viruses Indirect immunofluorescence analysis of viral proteins in infected human cells Quantitative analysis of apoptosis during HSV infection Temporal analysis of productive viral protein synthesis by denaturing gel and immunoblotting methods M. Tevfik Dorak, Ph.D - Genotyping by real-time PCR - Genotyping by high-resolution melting analysis  - Genotyping by fragment analysis - Genotyping by sequence analysis Grant Gallagher, Ph.D Flow Cytometry and FACS analysis Construction and analysis of vectors for promoter reporter activity quantitative real-time PCR for human mRNA species mammalian expression of human proteins and cell signalling preparation and culture of human primary and transformed cells and cell-lines Scott Gygax, Ph.D Antimicrobial Susceptibility Testing (Includes clinical bacterial and yeast isolation and CLSI AST methods) qRT-PCR methods   Kathy Iacono, Ph.D Pyrosequencing and Bio-Plex analyses of DNA Molecular Cloning methods Conventional and RT-PCR methods The use of robotics in molecular diagnostic analysis Joe Nickels, Ph.D Culture methods for mammalian, bacterial, and yeast cell growth Lenti virus infection methods and their use in construction of stable cell lines Molecular methods for DNA mutagenesis Molecular methods for promoter activity analysis Molecular methods for protein expression and purification Jason Trama, Ph.D Gene Expression by Quantitative RT-PCR Cytokine Expression by ELISA Protein-Protein Interactions by Immunoprecipitation Protein Expression by SDS-PAGE (1D and 2D) and Western Blotting Clinical Sample Processing Cell Culture John.P. Vermitsky, Ph.D Microbial mutagenesis and gene disruption methods Western blotting methods for protein analysis