Unknown sequence amplification: application to in vitro genome walking in Chlamydia trachomatis L2.
A recently described technique, 'Chemical Genetics' unknown sequence amplification method, which requires only one specific oligonucleotide, has broadened the applicability of the polymerase chain reaction to DNA of unknown sequence. We have adapted this technique to the study of the genome of Chlamydia trachomatis, an obligate intracellular bacterium, and describe modifications that significantly improve the utility of this approach. These techniques allow for rapid genomic analysis entirely in vitro, using DNA of limited quantity or purity.
ICI Diagnostics, Rudheath, Northwich, Cheshire, UK.
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D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D002692:Chlamydia trachomatis / Q000235:genetics*; D002874:Chromosome Mapping; D003545:Cysteine / Q000235:genetics; D004247:DNA / Q000235:genetics; D005784:Gene Amplification; D015698:Genomic Library; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D012335:RNA, Ribosomal; D004247:DNA; D003545:Cysteine