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In: Computers and Technology

Submitted By wine32
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Current Applications of Genetic Technology in Predisposition Testing and Microsatellite Instability Assays
By Marsha L. Frazier, Li-Kuo Su, Christopher I. Amos, Patrick M. Lynch
From the Departments of Epidemiology, Gastrointestinal Medical Oncology and Digestive Diseases, and Molecular and Cellular Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, TX.
Address reprint requests to Marsha L. Frazier, MD, Department of Epidemiology, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX 77030; email mlfrazier@notes .mdacc.tmc.edu.
INTRODUCTION
IT IS POSSIBLE TO test selected subjects for germline mutations in genes causing familial adenomatous polyposis (FAP),1 hereditary nonpolyposis colorectal cancer(HNPCC),2-8 Peutz-Jeghers syndrome,9,10 and juvenile polyposis.11-13 Because the genes that are mutated in familial colorectal cancer syndromes can be mutated at a variety of different locations, assays for mutation detection are not simple. Many different approaches to mutation detection have been described in the literature, some of which are also described here. Specific strategies for testing are also discussed.
THE BASICS
Isolation of DNA and Polymerase Chain Reaction (PCR)
DNA or RNA for genetic testing is almost always isolated from peripheral-blood leukocytes. This requires that the blood be drawn in tubes containing some sort of anticoagulant. The preferred anticoagulants are either citrate or EDTA. The cells are lysed followed by removal of the other cellular components and precipitation of the DNA or RNA in ethanol. One of the drawbacks of this approach is that the blood must be rapidly transported to the laboratory where the testing will be performed before the nucleic acids begin to degrade. Recent developments in filter paper technology show promise in obviating this problem. Special filter papers

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