Figure 1.

LDI-PCR based method to detect the activity of a hot L1. (a) Schematic showing mobility of hot L1 from the TTC28 locus upon activation. (b) LDI-PCR to detect 3′ transduction arising from the L1 at TTC28: Schematic representation of a hypothetical TTC28 specific L1 retrotransposition including transduction of 3′ flanking region or the “unique tag” (=region between the canonical polyadenylation signal and an alternative polyadenylation signal downstream), into an unknown target locus. NsiI produces restriction fragments of two different sizes that are self-ligated to form a circular template. Upon LDI-PCR, an inverse primer pair directed at the unique tag produces a native product and an insertion-specific target product. In addition to NsiI, two further restriction enzymes (PstI and SacI) and primer pairs (not depicted here) were used; see Materials and Methods for details.