+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Information on DNA conformation derived from the Ferguson plot of DNA fragments of up to 9 kb in size, using polyacrylamide gel electrophoresis in a discontinuous buffer system



Information on DNA conformation derived from the Ferguson plot of DNA fragments of up to 9 kb in size, using polyacrylamide gel electrophoresis in a discontinuous buffer system



Electrophoresis 12(4): 241-246



The Ferguson plot in polyacrylamide gel electrophoresis (PAGE) (15% CDATD, moving boundary electrophoresis buffer system operative at pH 8.9, 4.degree.C, 8 mA/cm2 of gel) of DNA fragments up to 9.4 kb in size was found to exhibit a linear segment at polyacrylamide concentrations starting at 3%T and undergoing a gradual transition into a concave segment at higher gel concentrations, confirming previous findings by Stellwagen [1]. The larger the DNA, and the higher the gel concentration, the less extended the linear and the more extended the concave segment of the plot. The lowest %T of the linear range for DNA in polyacrylamide remains unknown since mobilities at nongelling concentrations below 3%T have not as yet been measured. As previously suggested [1, 2], the transition from the linear to the concave segment corresponds to that from the randomly oriented DNA to the anisotropically stretched, "reptating" DNA. For a DNA of 9.4 kb in size, the end of the linear range of the Ferguson plot can be extended from 3.5 to 5%T when 15% DATD rather than 2.5% Bis is used to crosslink the polyacrylamide. Increasing the temperature of PAGE from 4.degree.C to 25 and 50.degree.C widens the linear segment progressively, indicating an increasingly random orientation with rising temperature. When current density is increased from 8 to 40 mA/cm2, the concave curvature f the Ferguson plot of DNA 1 to 9.4 kb in size decreases, suggesting a transition from a "reptating" to a randomly distributed molecule, due to increased Joule heat. Increasing the ionic strength from 0.01 to 0.04 M similarly appears to lead to a relative linearization of the Ferguson plot, suggesting a similar effect of Joule heat. Prestaining of DNA with ethidium bromide results in a decrease of both its size and its free mobility. This effect is exacerbated as the size of DNA increases from 0.1 to 9.4 kb.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 040439256

Download citation: RISBibTeXText

PMID: 1712721

DOI: 10.1002/elps.1150120403


Related references

Discontinuous buffer system for polyacrylamide and agarose gel electrophoresis of DNA fragments. Electrophoresis 12(4): 233-240, 1991

An apparatus for thin layer vertical polyacrylamide gel electrophoresis with discontinuous buffer and gel system. Analytical Biochemistry 63(1): 241-248, 1975

New types of large-pore polyacrylamide-agarose mixed-bed matrices for DNA electrophoresis: pore size estimation from Ferguson plots of DNA fragments. Electrophoresis 16(8): 1337-1344, 1995

Program in BASIC for Ferguson plot analysis, using a personal computer: application to gel electrophoresis in a continuous buffer. Electrophoresis 10(3): 199-214, 1989

A thin-layer multistrip polyacrylamide gel electrophoresis apparatus for Ferguson plot analysis at the submicrogram load level. Electrophoresis 10(10): 726-729, 1989

Some aspects of the use of continuous and discontinuous buffer systems in polyacrylamide gel electrophoresis. Anal Biochem 11(2): 219-223, 1965

Discontinuous buffer systems for analytical and preparative electrophoresis of enzymes on polyacrylamide gel. Analytical Biochemistry 45(1): 68-85, 1972

Simultaneous Ferguson plot analysis, using electrophoresis on a single agarose pore gradient gel, of DNA fragments contained in a mixture. Electrophoresis 12(12): 997-1004, 1991

A convex ferguson plot of polystyrene particles in electrophoresis on 0.25 to 2.0 percent polyacrylamide 30 percent bis crosslinked. Electrophoresis 8(10): 471-476, 1987

Particle and gel fiber properties derived from the mobility in gel electrophoresis the dialectics of the ferguson plot. Schafer-Nielsen, C (Ed ) Electrophoresis '88; Sixth Meeting Of The International Electrophoresis Society, Copenhagen, Denmark, July 4-7, 502p Vch Publishers, Inc : New York, New York, Usa; Protein Laboratory, University Of Copenhagen: Copenhagen, Denmark (Dist By Vch Verlagsgesellschaft Mbh: Weinheim, West Germany) Illus Paper 28-48, 1988

A comparison of commonly used discontinuous and continuous buffer systems for electrophoresis in sodium dodecyl sulfate-containing polyacrylamide gels. Electrophoresis 1(3-4): 180-186, 1980

Estimation of polyacrylamide gel pore size from Ferguson plots of linear DNA fragments. II. Comparison of gels with different crosslinker concentrations, added agarose and added linear polyacrylamide. Electrophoresis 12(9): 612-619, 1991

Estimation of polyacrylamide gel pore size from Ferguson plots of normal and anomalously migrating DNA fragments. I. Gels containing 3% N,N'-methylenebisacrylamide. Electrophoresis 12(4): 253-263, 1991

System for simplified discontinuous-gradient polyacrylamide-gel electrophoresis. Clinical Chemistry 23(3): 590-592, 1977

A vertical flat-bed discontinuous electrophoresis system in polyacrylamide gel. Analytical Biochemistry 16(3): 457-465, 1966