Automated capture and on-column detection of biotinylated DNA on a disposable solid support.
This work comprises the development of a technique for the capture of single-stranded DNA on a solid support combined with in situ quantification. The capture is based on the strong and selective interaction between biotinylated DNA and streptavidin-coated agarose beads. Sequential Injection in the lab-on-valve format allows for automated manipulation of all components including the building and disposal of bead columns. Detection was accomplished using the OliGreen fluorescent dye and optimization of the assay achieved a limit of detection of 111 pg ssDNA, with a total assay time of roughly 2.5 min per sample.
Decuir
Michael
M
Department of Chemistry, University of Washington, Box 351700, Seattle, WA 98195-1700, USA.
Lähdesmäki
Ilkka
I
Carroll
Andrea D
AD
Ruzicka
Jaromir
J
D001322:Autoanalysis / Q000295:instrumentation / Q000379:methods*; D001710:Biotin; D002138:Calibration; D004277:DNA, Single-Stranded / Q000032:analysis*; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods; D005456:Fluorescent Dyes; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D053758:Nanoparticles; D009691:Nucleic Acid Denaturation
D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D001710:Biotin