Localized transfection with magnetic beads coated with PCR products and other nucleic acids.
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.
Santori
Maria Isabel
MI
EMBL-CRG Systems Biology Programme, Centre for Genomic Regulation, Passeig Maritim 37-49, E-08003 Barcelona, Spain.
Gonzalez
Cayetano
C
Serrano
Luis
L
Isalan
Mark
M
D002460:Cell Line; D006801:Humans; D008280:Magnetics; D009696:Nucleic Acids / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000295:instrumentation / Q000379:methods*
D009696:Nucleic Acids