A novel functional micropipette tip for DNA purification, called DNAtip, is described based on a novel coating process. The functional layer is prepared by an internal coating with silica particles supported into a polyvinyl alcohol (PVA) film. The coating layer is stabilized by simultaneous radiation-induced crosslinking to avoid its dissolution. Three types of silica particles of different diameters: micron, sub-micron, and nano sizes are studied. In addition, pH of the coating solution and concentration of the silica particles were optimized. Tips coated with fumed silica showed the highest absorption capacity of a plasmid DNA with an acceptable purity. Different linear dsDNA fragments were used for evaluating the absorption capacity and size discrimination of fragments. The recovery in the range of 700 ng and 200 ng per tip was measured for dsDNA fragments in the range of 400 bp to 2 kbp and 50 bp to 500 bp respectively. DNA PCR fragments contaminated with BSA were successfully purified with sufficient quality to be sequenced or enzymatically digested. This disposable tip device, useful for plasmids or DNA fragments purification, is a novel lab-on-a-tip device, which can effectively improve automate genetic engineering procedures.
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This work was supported by grants from Universidad Nacional de Quilmes and Agencia Nacional De Promoción Científica y Tecnológica (ANPCyT) (PICT 2010-0336 and PICT 2014-3771). CYG and LJM thank CONICET for their fellowships. MLS, MR and MG are researchers from CONICET.
The authors declare no conflict of interest.
Neither ethical approval nor informed consent was required for this study.
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Sánchez, M.L., Gimenez, C., Martínez, L.J. et al. Disposable Micropipette Tip for Purifying DNA Fragments. Biotechnol Bioproc E 25, 215–223 (2020). https://doi.org/10.1007/s12257-019-0141-3
- DNA purification
- DNA purity
- disposable tips