Lariat-dependent nested PCR for flanking sequence determination.
Methods detailed in this chapter relate to the use of Lariat-dependent Nested (LaNe) PCR to characterize unknown RNA or DNA sequence flanking known regions. A multitude of approaches designed to determine flanking sequences have been described in the literature. Variously, problems related to these approaches include lack of resolution or failure, depending on experimental context, and complex handling. LaNe-based methods are designed to harness "two-sided" gene-specific PCR with the option of nesting but without the requirement for inefficient and involved enzyme preprocessing steps.
Genetic Epidemiology Laboratory, Department of Pathology, The University of Melbourne, Parkville, Victoria, Australia. djp@unimelb.edu.au
Daniel J
DJ
Park
D001483:Base Sequence; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000235:genetics
D017931:DNA Primers; D012313:RNA; D004247:DNA