Detection of -mediated Transposition by Splinkerette PCR in Transgenic Lines of .

Splinkerette PCR (spPCR) is a newly developed and efficient method to ascertain and characterize genomic insertion sites of transgenes. The method described in this protocol was successfully applied to confirm transposon-mediated integration of transgenes into chromosomes of the parasitic nematode . This work is described in detail in Shao . (2012) and presented here in a simplified diagram (Figure 1). Using this method, chromosomal loci of integration were determined based on target site and 5'- and 3' flanking sequences. Therefore, spPCR can be a useful method to confirm integrative transgenesis in functional genomic studies of parasitic nematodes. Potter and Luo (2010) contains a protocol for use of spPCR to detect and map transposon-mediated chromosomal integrations in , and was the source of our method for . The splinkerette- and specific oligos described in that reference could be used without modification in . For interested readers, a general review of the biology of parasitic nematodes in the genus may be found in Viney and Lok (2007), and a methods-based article on as an experimental model, with information on transgenesis, may be found in Lok (2007).