[10] High-speed automated DNA sequencing in ultrathin slab gels

Cross-linked polyacrylamide gel electrophoresis of single-stranded DNA for microfabricated genomic analysis systems

“…Human identification genotyping requires slightly higher electrophoresis resolution but nevertheless stands to benefit from the miniaturization and automation associated with microfabricated systems. Current human genotyping (e.g., forensic and paternity testing) relies on using PCR to amplify a modest number of identity loci (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) followed by gel electrophoresis to detect allelic series variations which typically occur at 2-4 base intervals. Single nucleotide polymorphism (SNP) assays are another area receiving increased attention as a vehicle to identify the genetic basis for many diseases.…”

“…The relatively large dimensions of macroscale slab gels, however, make them especially susceptible to nonuniformities in DNA migration velocity arising as a consequence of Joule heating at high electric field strengths [7]. These adverse effects, however, can be minimized through careful temperature control and the use of ultrathin slab gels [8].…”

DNA Technologies: Sequencing Technology

“…Several methods have been developed to sequence DNA using fluorescent labels (ANSOR- GE et al, 1986;SMITH et al, 1986;PROBER et al, 1987;BRUMBAUGH et al, 1988). Commercialized instruments employ one or more of the following methods for automated sequencing: four distinct dye-labeled primers with nonfluorescent terminators per DNA sample; one dye-labeled primer with non-fluorescent terminators per DNA sample; and one non-fluorescent primer with four distinct fluorescent terminators per DNA sample (see Sect.…”

“…The first reports of automation of DNA sequencing occurred in the mid-1980s due to novel techniques to fluorescently label DNA (SMITH et al, 1986;ANSORGE et al, 1986ANSORGE et al, ,1987PROBER et al, 1987;BRUMBAUGH et al, 1988;KAMBARA et al, 1988;MIDDENDORF et al, 1988).This automation, in conjunction with the commencement of the human genome initiative (DELISI, 1988), spurred the explosion in genomics research that is in existence today. DNA sequencing technology is now only one tool, albeit a very important and dynamic one, in the genomics toolbox along with other tools such as DNA array and lab-on-a-chip technologies as well as automated protein analysis.…”

Sequencing Technology

“…The use of high voltage gradients (see Sect. 5.1.4) in combination with 203 either ultrathin slab gels (BRUMLEY and SMITH, 1991;KOSTICHKA et al, 1992;CARNINCI et al, 1995;SMITH et al, 1996;YAGER et al, 1997) or capillary electrophoresis has significantly reduced run times, but at the expense of read length (LUCKEY and SMITH, 1993a;YAN et al, 1996). Efforts to increase read length for capillary DNA sequencing are under development (KLEPARNIK et al, 1996;CARRILHO et al, 1996;SALAS-SOLANO et al, 1998b).…”