A RELIABLE AMPLIFICATION TECHNIQUE FOR THE CHARACTERIZATION OF GENOMIC DNA-SEQUENCES FLANKING INSERTION SEQUENCES

Authors
PRODHOM G LAGIER B PELICIC V HANCE AJ GICQUEL B GUILHOT C
Citation
G. Prodhom et al., A RELIABLE AMPLIFICATION TECHNIQUE FOR THE CHARACTERIZATION OF GENOMIC DNA-SEQUENCES FLANKING INSERTION SEQUENCES, FEMS microbiology letters, 158(1), 1998, pp. 75-81
Citations number
11
Categorie Soggetti
Microbiology
Journal title
FEMS microbiology letters → ACNP
ISSN journal
03781097
Volume
158
Issue
1
Year of publication
1998
Pages
75 - 81
Database
ISI
SICI code
0378-1097(1998)158:1<75:ARATFT>2.0.ZU;2-G
Abstract
A simple and efficient ligation-mediated PCR (LMPCR) is described for amplifying DNA adjacent to known sequences. The method uses one primer specific for the known sequence and a second specific for a synthetic linker ligated to restricted genomic DNA. Perkin-Elmer AmpliTaq Gold polymerase is used to minimize non-specific primer annealing and ampli fication. This LMPCR method was successfully applied to isolate DNA se quences flanking mobile elements present in mycobacterial mutants gene rated by transposon mutagenesis. (C) 1998 Published by Elsevier Scienc e B.V. All rights reserved.