Gene identification by arrested primer extension.
This paper reports a novel method for the identification of nucleic acid target sequences when these targets have high sequence identity. Homologous genes are currently identified by sequencing. We hypothesize that by primer extension in the presence of selected nucleotides, genes with similar sequence can be identified by the length of the extension products on gel electrophoresis. This simple procedure eliminates the much-demanding process of sequencing. We term this process Arrested Primer Extension (APE). As a demonstration of the feasibility of this method, we have used APE to speciate a known set of cultured mycobacteria. There should be many other applications of this method.
Sandhu
G S
GS
Department of Biochemistry and Molecular Biology, Mayo Clinic and Foundation, Rochester, MN 55905.
Kline
B C
BC
Bolander
M E
ME
Sarkar
G
G
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics; D003854:Deoxyribonucleotides; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D003854:Deoxyribonucleotides; D009838:Oligodeoxyribonucleotides; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S