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Cystic Fibrosis: Methods and Protocols consolidates cutting-edge in vitro, cellular, and whole animal laboratory protocols into an indispensable resource. From electrophysiology and cell biology, to animal models and gene therapy, this comprehensive set of methods provides the step-by-step instructions needed for investigators to incorporate new approaches into their research programs. Specific protocols describe new techniques for diagnosis, in vitro methods for the expression and functional analysis of CFTR, novel biochemical and cellular systems to determine how mutations subvert CFTR function, and in vivo protocols to examine how CFTR dysfunction produce multisystem pathology in both human and animal models.
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M E T H O D S I N M O L E C U L A R M E D I C I N E TM Cystic Fibrosis Methods and Protocols Edited by William R. Skach, MD Humana Press CFTR Mutation Detection by mHET Analysis 3 1 CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel Julian Zielenski, Isabel Aznarez, Tuncer Onay, John Tzounzouris, Danuta Markiewicz, and Lap-Chee Tsui 1. Introduction Mutation detection in an integral part of disease diagnosis and patient study. For most Mendelian diseases, multiple mutations may be found in a single gene among a patient population. The type of mutations may vary from large deletions to single-base-pair (bp) substitutions, and different diseases may have different predominant types. For example, large deletions are often found in Duchenne muscular dystrophy (1) and truncation mutation is the predominant type in BRCA1-associated breast cancer (2). Therefore, different mutation detection strategies are required for different diseases. Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gen