Use of high-molecular-mass polyacrylamides as matrices for microchip electrophoresis of DNA fragments.
DNA fragment analysis requires the use of polymer solutions as sieving matrices. Generally, such matrices are constituted of high-molar-weight polymers employed at a concentration higher than their entanglement threshold concentration. These polymer solutions are highly viscous and difficult to use in the narrow channels of a microchip. Ultralarge polyacrylamides synthesized via a nonconventional method, being the low-temperature plasma-induced polymerization (PIP), were used as DNA sieving matrices for microchip electrophoresis. The distinctive features of these polymers (ultralarge molecular mass and linearity) allow their use at a dilute concentration. Dilute PIP polyacrylamides revealed a constant value of resolution in a broad range of DNA fragment sizes (123 bp-1353 bp), thus proving to be effective in common genotyping applications. Moreover, the low viscosity of the dilute solutions enable it to be easier and faster in filling the channel between runs, thus enhancing the throughput of the microchip devices.
Cretich
Marina
M
Istituto di Chimica del Riconoscimento Molecolare, C.N.R., Milano, Italy. marina.cretich@icrm.cnr.it
Chiari
Marcella
M
Rech
Ivan
I
Cova
Sergio
S
D000180:Acrylic Resins; D020029:Base Pairing; D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D007202:Indicators and Reagents; D008904:Miniaturization / Q000295:instrumentation / Q000379:methods; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D011089:Polydeoxyribonucleotides / Q000737:chemistry* / Q000302:isolation & purification
D000180:Acrylic Resins; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D011089:Polydeoxyribonucleotides; C016680:polyacrylamide gels; D004247:DNA