Figure 3

PCR based assay of nuclear matrix associated DNA from Giardia. Nuclear matrix dependent (A;Lane 1) and independent DNA (A;Lane 2) was extracted from nuclear matrix (see methods) according to [20 and used for PCR reactions with predicted S/MAR primers(B). Lane M is the DNA marker (NEB); lanes 1-30 are PCR products of which odd number lanes (1,3,5,7.....27,29) even numberedare nuclear lanes (2,4,6...26,28,30) are nuclear matrix from the Giardia genomic DNA as a positive control(C). The PCRs are loaded in the following order: GlSMAR3, GlSMAR7, GlSMAR10, GlSMAR11, GlSMAR16, GlSMAR20, GlSMAR66, GlSMAR20, GlSMAR22, GlSMAR26-1, GlSMAR26-2, GlSMAR39, GlSMAR42, GlSMAR51, GlSMAR55, GlSMAR58, in both panel B and C. A non-S/MAR sequence was used as a control (PanelD). Amplification of multiple bands was seen in the matrix independent fraction (loop-fraction) lane 2 and no amplification was seen in the matrix dependent fraction.