Basic Techniques in Molecular Pathology

Abstract

A critical step in any molecular analysis is in the determination and understanding of the material being selected. There are a variety of sample types than can be used, including; cell culture, body fluids, and both fresh and fixed tissue. All of these have various advantages and disadvantages that must be carefully considered. Many different types of molecular techniques are available for use in both a research and clinical setting. These range from simple, fast assays that interrogate a single area to large, complex assays that can sequence the entire genome. While technology is rapidly evolving, some of the more common technologies used to interrogate a single loci, gene, or RNA transcript include quantitative real-time polymerase chain reaction (qPCR), mutation-specific polymerase chain reaction, and in situ hybridization (ISH). Two technologies that are commonly used to analyze a larger/broader area include microarrays and sequencing. Currently, massively parallel sequencing (MPS) is being used for a variety of large-scale analysis. These range from whole genome sequencing (WGS), expression analysis using RNA-sequencing, and determining where certain transcription factors bind on the genome using chromatin immunoprecipitation sequencing. Regardless, a tremendous amount of data can be generated and often a single person does not have all of the desired expertise to effectively handle all of the requirements necessary to properly generate and interpret this amount of data. Therefore, collaboration with experts in other fields is often necessary to generate, analyze, and interpret all of the data.