Spatial and temporal depletion of ions from noncrosslinked denaturing polyacrylamide in capillary electrophoresis

Figeys, Daniel; Renborg, Annika; Dovic̀hi, Norman J.

doi:10.1002/elps.11501501216

Cited by 20 publications

(16 citation statements)

References 32 publications

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“…[16], Baba et al [14], and Figeys et al [18]. However, the effects of polymer persistence length ( L e .…”

Section: Introductionmentioning

confidence: 98%

“…Several different types of polymers are useful for such separations. To date, researchers have used at least twelve different water-soluble polymers as DNA separation media for CE, including methyl cellulose , hydroxyethylcellulose (HEC) , hydroxypropylcellulose (HPC) [14], hydroxypropylmethylcellulose [14-181, polyethylene glycol [15, 191, galactomannan [19], and polyvinyl alcohol [16, 19,201, as well as liquified agarose [21], linear polyacryloylaminoethoxyethanol [22], and linear polyacrylamide (PAA) [ 18,20,[23][24][25][26][27].…”

Section: Introductionmentioning

confidence: 99%

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The effects of polymer properties on DNA separations by capillary electrophoresis in uncross‐linked polymer solutions

1996

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Low-viscosity, aqueous solutions of hydrophilic linear polymers have been shown to be useful for the separation of DNA restriction fragments by capillary electrophoresis (CE). However, the choice of polymer type, size, and concentration remains largely empirical, because the mechanism of high-field electrophoretic DNA separations in polymer solutions is not well understood. To assist in elucidating the mechanism of DNA separation, we experimentally investigated the effects of polymer properties such as stiffness (persistence length), average molecular mass, polydispersity, and hydrophilicity on the separation of DNA ranging from 72 bp to 23 kbp. This was accomplished by comparing the results of DNA separations obtained by counter-migration CE in dilute and semidilute solutions of linear polyacrylamide (PAA), hydroxyethyl-cellulose (HEC), and hydroxypropylcellulose (HPC) polymers of several different average molecular masses.

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“…[16], Baba et al [14], and Figeys et al [18]. However, the effects of polymer persistence length ( L e .…”

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

The effects of polymer properties on DNA separations by capillary electrophoresis in uncross‐linked polymer solutions

1996

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“…There was an optimum period in the life of polymer when the resistive zone formed less readily. They also reported a negative effect of the template presence [9][10][11], explored the possibility of multiple use of capillary polymer [11], and studied the effect of buffer composition [12].…”

Section: Introductionmentioning

confidence: 99%

“…Most relevant to our experience is the series of works from The University of Alberta [9][10][11][12] that investigated processes in linear polyacrylamide during the electrophoresis in precoated capillaries. The resistive zone in their capillaries was localized in the first few centimeters at the injection end.…”

Section: Introductionmentioning

confidence: 99%

Formation of a resistive region at the anode end in DNA capillary electrophoresis

et al. 2003

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We have studied the formation of a resistive region in the capillary during DNA separation. This effect is caused by an unequal change in the mobilities of cations and anions at the interface between the running buffer solution and the capillary. We studied the motion of the resistive region boundary by sequential removal of portions of the affected capillary end. We found that in the process of developing the resistive region the distribution of the electric fields in the capillary changes from uniform to extremely nonuniform, with a very high field (above 1 MV/cm) in the resistive region and a reduced field (80 V/cm) in the rest of the capillary. Though theoretically a resistive region may appear either at the anode (detection) or the cathode (injection) end of the capillary, all previous publications report the formation of the resistive region at the cathode side. In our experiments, however, the anomalous region is formed at the anode. Thus, the separated DNA peaks move towards the slowly progressing resistive region. Our results indicate that the DNA is stopped at the boundary and does not enter the region. When the resistive region is clipped off the peak motion resumes. This suggests that there exists a potential barrier at the resistive layer boundary that prevents the drift of the peaks towards the anode. The formation of the resistive region interferes with a normal separation process causing a gradual decrease of the capillary current and the deceleration and eventual quenching of the peak motion. For the ABI chemistry, we experimented with adding polymers to the electrode buffer to equate the transference numbers for anions and cations, and found the conditions at which this effect is completely eliminated.

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“…However, CGE still had some features that were not compatible with a high-throughput regime. Among them were the short column lifetime as well as injection-related problems [29,30].…”

Section: Competing Technologies For Highthroughput Dna Sequencingmentioning

confidence: 99%

DNA sequencing by capillary array electrophoresis and microfabricated array systems

Carrilho

2000

Electrophoresis

113

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To comply with the current needs for high‐speed DNA sequencing analysis, several instruments and innovative technologies have been introduced by several groups in recent years. This review article discusses and compares the issues regarding high‐throughput DNA sequencing by electrophoretic methods in miniaturized systems, such as capillaries, capillary arrays, and microchannels. Initially, general features of several capillary array designs (including commercial ones) will be considered, followed by similar analyses with microfabricated array electrophoretic devices and how they can contribute to the success of large sequencing projects.

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Spatial and temporal depletion of ions from noncrosslinked denaturing polyacrylamide in capillary electrophoresis

Cited by 20 publications

References 32 publications

The effects of polymer properties on DNA separations by capillary electrophoresis in uncross‐linked polymer solutions

The effects of polymer properties on DNA separations by capillary electrophoresis in uncross‐linked polymer solutions

Formation of a resistive region at the anode end in DNA capillary electrophoresis

DNA sequencing by capillary array electrophoresis and microfabricated array systems

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