Table 3. Comparison of in vitro with in silico results from PCR-RFLP-ITS2 assay.

Species	Agarose gel fragment sizes (bp)		Bioinformatic prediction of fragment sizes (bp)
	PCR product	PCR-RFLP	PCR product	PCR-RFLP
An. neomaculipalpus	500	259, 165, 111	452	255, 136, 61
An. apicimacula lineage C (Caribbean)	481	300, 150, 70	481	290, 130, 61
An. apicimacula lineage P (Pacific)	480	390, 70	480	372, 61, 47
An. punctimacula s.s.	397	334, 81	393	317, 76
An. calderoni	394	Uncut	401	Uncut
An. malefactor	399	Uncut; 378, 313, 113	401; 396	Uncut; 326, 70
An. mattogrossensis	505	230, 80	464*	213, 190, 61
An. peryassui	500	300, 96	464*	287, 116, 61

* An ITS2 variant of 463 bp was found that yielded 286, 116 and 61 bp in the in silico restriction analysis.

PCR: Polymerase Chain Reaction; RFLP: Restriction Fragment Length Polymorphisms; (bp): base pairs.