The ubiquity of pollen allows for it to serve as a forensic marker in criminal cases where temporal and spatial origins are of importance. Current palynological analyses of pollen focuses on the morphology of grains, with few researchers aimed at exploring its genetic capabilities to serve as a biomarker. For pollen to be subject to DNA analysis, a sufficient yield must first be obtained during collection. Current collection methods are limited and so this research focused on determining the best mode of collection of Ruellia simplex pollen by comparing tape lifting with the use of the Venturi Vacuum Device (VVD) on two collection fabrics of different textures: 100% woven cotton t-shirt and 100% twill cotton denim. DNA from the pollen was then extracted and quantified to determine which collection method produced the highest quantity of DNA. The DNA was further analyzed via amplification using trnL and rbcL primers and fragment analysis to determine the level of taxonomic resolution achieved for each collection method. Findings suggest that tape lifting may be the better method of the two for pollen collection, although reproducibility and quantity of DNA extracted was inconsistent for both collection methods. There is therefore a need to optimize DNA extraction from Ruellia simplex pollen grains to counteract this low yield and improve the potential use of pollen as a forensic tool. Both trnL and rbcL primers successfully amplified DNA specific to plant material and their respective amplicon sizes coincided with that of Ruellia simplex pollen. Therefore, this confirms the potential of pollen as a biomarker in crime scene investigations, but optimization and reproducibility are key.
