Lariat-Dependent Nested PCR for Flanking Sequence Determination

Abstract

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.