Electrokinetic injection of DNA from gel micropads: basis for coupling polony technology with CE separation.
We propose a novel method for electrokinetic injection of DNA samples into capillaries from nanoliter gel micropads, deposited on glass slides, which are coated with electroconducting film. Theoretical and experimental proof is presented for the proposed method. The method allows efficient and highly precise injection without physical contact between the gel pad and the capillary. Read length of more than 700 bp at Q20 has been reproducibly demonstrated in fused-silica capillaries using the proposed injection technique. Based on the obtained results we discuss a novel DNA sequencing system which combines DNA amplification and cycle sequencing in arrays of subnanoliter gel micropads and high-throughput electrophoretic separation in monolith multicapillary arrays.
Kosobokova
Olga
O
Department of Electrical and Computer Engineering, State University of New York, Stony Brook, NY 11794-2350, USA. okey@ece.sunysb.edu
Gavrilov
Dmitri N
DN
Khozikov
Vyacheslav
V
Stepukhovich
Andrey
A
Tsupryk
Andriy
A
Pan'kov
Sergey
S
Somova
Olga
O
Abanshin
Nikolai
N
Gudkov
Georgiy
G
Tcherevishnik
Marina
M
Gorfinkel
Vera
V
D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D005782:Gels / Q000737:chemistry; D007202:Indicators and Reagents; D008956:Models, Chemical; D036103:Nanotechnology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet
D005782:Gels; D007202:Indicators and Reagents; D004247:DNA