Localized transfection with magnetic beads coated with PCR products and other nucleic acids |
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Authors: | Santori Maria Isabel Gonzalez Cayetano Serrano Luis Isalan Mark |
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Affiliation: | EMBL-CRG Systems Biology Programme, Centre for Genomic Regulation, Passeig Maritim 37-49, E-08003 Barcelona, Spain. |
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Abstract: | The bead transfection method involves binding nucleic acids onto 3-microm-diameter paramagnetic beads, treating the beads with transfection reagent, and using them as scaffolds to direct transfection to individual cells or regions in a population. Typically, PCR products are used because they can be conveniently generated using biotinylated primers and can introduce site-directed mutations, without the need for cloning or plasmid purification. However, the method can be adapted to transfect plasmid DNA or RNA. The magnetic properties of the beads allows magnets to direct the loci of transfection in cell culture; magnetic arrays are built in cell culture chambers to allow multiple parallel transfections on the same microscope coverslip. The PCR reaction and transfection can be carried out in 1 d, and transfection results can be viewed in 24-48 h. |
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