The limiting mobility of DNA sequencing fragments for both cross‐linked and noncross‐linked polymers in capillary electrophoresis: DNA sequencing at 1200 V cm<sup>−1</sup>

Yan, Juying; Best, Norine; Zhang, Jianzhong; Ren, Hongji; Jiang, Rong; Hou, Joan Y.; Dovic̀hi, Norman J.

doi:10.1002/elps.1150170611

Cited by 43 publications

(25 citation statements)

References 34 publications

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“…At relatively low electric fields, one can observe two regimes with m z N bl 21 and m z N bl 0 , respectively. The same kind of scaling was reported by Slater and Drouin [70], Yan et al [71] and Pluen [72] for ssDNA in polyacrylamide gels. Viovy [73] showed that the theoretical result of Eq.…”

Section: Discussionsupporting

confidence: 72%

Sieving mechanisms in polymeric matrices

2003

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A critical review of the existing theoretical models and experimental evidences for sieving mechanisms during separation of macromolecules, paying particular attention to capillary electrophoresis applications is presented. Gel models (Ogston and reptation) have been successfully applied to highly entangled polymer solutions, where fast and efficient separations can occur. In order to account for the DNA/polymers collision-interaction mechanisms during separation in dilute solutions - characterized by a poorer resolution -, approximated analytical models have been developed. An insight in the mechanism regulating the intermediate case of moderately entangled polymer solutions, for low fields and concentrations of small multiples of the overlap concentration c*, is given by the constraint release approach. This model proposes an upper limit of size separation, increasing with matrix concentration and molecular mass. Finally, the coupling between the reptative motion of the analytes and the effect of matrix constraint release very likely plays a fundamental role in the separation mechanism and requires therefore further and deeper investigation, both theoretically and experimentally.

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Section: Discussionsupporting

confidence: 72%

Sieving mechanisms in polymeric matrices

2003

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“…Available data in the literature usually deal with fields below 500 V/cm. In one work, the authors report results obtained at 1200 V/cm [20], however, their experiments were carried out in cross-linked polymer. No information is available on the zone behavior in the noncross-linked polymer at fields of the order that occur in the resistive region.…”

Section: Discussionmentioning

confidence: 91%

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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“…Our results are in contrast to findings from Dovichi©s group, who indeed reported a dependence of N* nearly inversely proportional to the electric field in 5% non-cross-linked polyacrylamide (see Figs. 10 and 11 of [40]). …”

Section: Comparison To Theoretical Predictionsmentioning

confidence: 90%

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Heller

1999

Electrophoresis

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We have studied the separation of single‐stranded and double‐stranded DNA in a matrix of entangled, linear poly‐N,N‐dimethylacrylamide. Our results give better insight into the mechanisms involved during separations in polymer solutions. The dependence of different parameters on DNA size, electric field, pore size and the polymer chain length are evaluated and compared to theoretical predictions. Striking differences between experimental data and predicted scaling laws are found. Our data should help to optimize DNA separation in capillary electrophoresis and to improve existing models for DNA separation in porous matrices.

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The limiting mobility of DNA sequencing fragments for both cross‐linked and noncross‐linked polymers in capillary electrophoresis: DNA sequencing at 1200 V cm⁻¹

Cited by 43 publications

References 34 publications

Sieving mechanisms in polymeric matrices

Sieving mechanisms in polymeric matrices

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

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

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The limiting mobility of DNA sequencing fragments for both cross‐linked and noncross‐linked polymers in capillary electrophoresis: DNA sequencing at 1200 V cm−1

Cited by 43 publications

References 34 publications

Sieving mechanisms in polymeric matrices

Sieving mechanisms in polymeric matrices

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

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

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Product

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The limiting mobility of DNA sequencing fragments for both cross‐linked and noncross‐linked polymers in capillary electrophoresis: DNA sequencing at 1200 V cm⁻¹