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Review
.2009 Jun;30 Suppl 1(Suppl 1):S196-202.
doi: 10.1002/elps.200900218.

DNA sequencing by CE

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Review

DNA sequencing by CE

Barry L Karger et al. Electrophoresis.2009 Jun.

Abstract

Sequencing of human and other genomes has been at the center of interest in the biomedical field over the past several decades and is now leading toward an era of personalized medicine. During this time, DNA-sequencing methods have evolved from the labor-intensive slab gel electrophoresis, through automated multiCE systems using fluorophore labeling with multispectral imaging, to the "next-generation" technologies of cyclic-array, hybridization based, nanopore and single molecule sequencing. Deciphering the genetic blueprint and follow-up confirmatory sequencing of Homo sapiens and other genomes were only possible with the advent of modern sequencing technologies that were a result of step-by-step advances with a contribution of academics, medical personnel and instrument companies. While next-generation sequencing is moving ahead at breakneck speed, the multicapillary electrophoretic systems played an essential role in the sequencing of the Human Genome, the foundation of the field of genomics. In this prospective, we wish to overview the role of CE in DNA sequencing based in part of several of our articles in this journal.

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Figures

Figure 1
Figure 1
Capillary gel electrophoresis separation of polydeoxyadenylic acids, p(dA)10–30,40–60. Capillary: 270 × 0.075 mm i.d.; running buffer: 0.1 M Tris/0.25 M borate/7 M urea, pH 8.3, with crosslinked polyacrylamide gel: 7.5% T and 3.3% C. The applied electric field was 400 V/cm. With permission from [8].
Figure 2
Figure 2
Comparison of the migration behavior of wild-type and mutated DNA strands oflucl gene. With permission from [21].
Figure 3
Figure 3
The effect of capillary temperature and denaturants on resolution of two compression regions (I and II) in the M13mp18 sequence from base 386 to 423. (A) 25°C, 3.5 M urea, 30% formamide; (B) 50°C, 7 M urea; (C) 50°C, 3.5 M urea, 30 % formamide. The sieving matrix was 3% w/v LPA in 50 mM Tris-TAPS. With permission from [30].
Figure 4
Figure 4
Sequencing to 1300 bases at 70°C using high molecular mass polymer matrix, optimized electric field and base calling software. With permission from [35].
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