Reliable determination of transposon insertion site in prokaryotes by direct sequencing.
We developed a method to identify the insertion sites of transposons in the chromosome of Salmonella using one step only. In this method, the Salmonella's genomic DNA is directly sequenced using a transposon internal primer. Reliable direct sequencing was achieved using high purity genomic DNA and an improved protocol for automated sequence machine. This note is intended to promote the use of direct sequencing, which we found to be reliable, efficient and inexpensive as compared to the other currently used methods.
Qimron
Udi
U
Department of Microbiology and Immunology, Faculty of Health Sciences, and the Cancer Research Center, Ben Gurion University of the Negev, Beer-Sheva 84105, Israel.
Madar
Neta
N
Ascarelli-Goell
Rivi
R
Elgrably-Weiss
Maya
M
Altuvia
Shoshy
S
Porgador
Angel
A
D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000032:analysis*; D005821:Genetic Techniques; D016680:Genome, Bacterial; D016254:Mutagenesis, Insertional / Q000379:methods*; D012486:Salmonella typhimurium / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004251:DNA Transposable Elements; D004269:DNA, Bacterial