PMID: 7580892Oct 1, 1994Paper

Parameters affecting the sensitivities of dideoxy fingerprinting and SSCP

PCR Methods and Applications
Q Liu, S S Sommer

Abstract

The goals of the present experiments are (1) to improve dideoxy fingerprinting (ddF) and (2) to utilize ddF as a tool to evaluate the relative merits of different conditions for single-strand conformation polymorphism (SSCP). ddF is performed by electrophoresing one dideoxy termination reaction through a nondenaturing gel. The ddF pattern can be divided into a "dideoxy component" and an "SSCP component". If dideoxy CTP (ddCTP) is utilized for the termination reaction of ddF, the dideoxy component is abnormal when an extra segment is produced by a sequence change that creates an extra C or when a segment is eliminated by a change of C to another base. All subsequent segments produced by the termination reaction constitute the SSCP component that contains the mutation in a nested series of ddCTP termination products. The SSCP component is informative if abnormal mobility is detected in one or more of the segments. Herein, we utilize 84 different single-base changes in the human factor IX gene to examine the effects of gel matrix, temperature, and different primers on the sensitivity of ddF. The effects of glycerol and cross-linker ratio were examined on fewer mutations. The following conclusions emerge: 1. The sensitivity of the ...Continue Reading

Citations

Jan 22, 1998·Mammalian Genome : Official Journal of the International Mammalian Genome Society·M DasJ Pelletier
Feb 19, 1998·International Journal for Parasitology·R B Gasser
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Aug 24, 1999·Veterinary Parasitology·R B Gasser
Nov 5, 1998·Breast Cancer Research and Treatment·L L ChuJ Pelletier
Feb 6, 1996·Proceedings of the National Academy of Sciences of the United States of America·J S KovachS S Sommer
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Nov 14, 2000·Physiological Genomics·M DasJ Pelletier
May 18, 1999·Analytical Biochemistry·Q LiuS S Sommer
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