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Optimization of random amplification of polymorphic DNA analysis for molecular subtyping of Escherichia coli O157
Author(s) -
Hopkins K.L.,
Hilton A.C.
Publication year - 2001
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2001.00871.x
Subject(s) - subtyping , rapd , escherichia coli , polymerase chain reaction , biology , genomic dna , dna , genetics , dna profiling , computational biology , microbiology and biotechnology , gene , population , computer science , genetic diversity , demography , sociology , programming language
K.L. HOPKINS AND A.C. HILTON. 2001. Random amplification of polymorphic DNA (RAPD) analysis using the polymerase chain reaction has proved to be a useful technique in the epidemiological investigation of micro‐organisms but may suffer from a lack of reproducibility in poorly optimized protocols. In this study a method of obtaining reproducible genomic fingerprints using RAPD analysis of Escherichia coli O157 is described. By systematic optimization of reaction conditions and selection of suitable primers, reproducible and discriminatory profiles could be obtained from all E. coli O157 strains tested. In addition, two other methods of obtaining reproducible profiles from E. coli O157 strains without the need to purify genomic DNA are described.