Field Inversion Gel Electrophoresis in Denaturing Polyacrylamide Gels

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1992 May 25

PMID 1598202

Authors

C Heller

S Beck

Affiliations

Soon will be listed here.

Abstract

The velocities of single stranded DNA molecules in denaturing polyacrylamide gels during symmetric and asymmetric field inversion were measured at different pulse times and gel concentrations. Under the conditions chosen in our study, pulse times as short as a few milliseconds lead to a retardation of DNA molecules larger than 400 bases. We found that a field inversion with an electric field in the forward direction of about double the strength of that applied in the backward direction is a good compromise between the degree of retardation, the temperature control requirements and the run time of the gel.

References

Gejman P, Sitaram N, Hsieh W, Gelernter J, Gershon E . The effects of field inversion electrophoresis on small DNA fragment mobility and its relevance to DNA polymorphism research. Appl Theor Electrophor. 1988; 1(1):29-34. View

Garoff H, Ansorge W . Improvements of DNA sequencing gels. Anal Biochem. 1981; 115(2):450-7. DOI: 10.1016/0003-2697(81)90031-2. View

Smith S, Heller C, Bustamante C . Model and computer simulations of the motion of DNA molecules during pulse field gel electrophoresis. Biochemistry. 1991; 30(21):5264-74. DOI: 10.1021/bi00235a021. View

Heller C, Pohl F . Field inversion gel electrophoresis with different pulse time ramps. Nucleic Acids Res. 1990; 18(21):6299-304. PMC: 332495. DOI: 10.1093/nar/18.21.6299. View

Heller C, Pohl F . A systematic study of field inversion gel electrophoresis. Nucleic Acids Res. 1989; 17(15):5989-6003. PMC: 318255. DOI: 10.1093/nar/17.15.5989. View

Slater G, Noolandi J . The biased reptation model of DNA gel electrophoresis: mobility vs molecular size and gel concentration. Biopolymers. 1989; 28(10):1781-91. DOI: 10.1002/bip.360281011. View

Birren B, Lai E, Hood L, Simon M . Pulsed field gel electrophoresis techniques for separating 1- to 50-kilobase DNA fragments. Anal Biochem. 1989; 177(2):282-6. DOI: 10.1016/0003-2697(89)90052-3. View

Lai E, Davi N, HOOD L . Effect of electric field switching on the electrophoretic mobility of single-stranded DNA molecules in polyacrylamide gels. Electrophoresis. 1989; 10(1):65-7. DOI: 10.1002/elps.1150100116. View

Smith S, Aldridge P, Callis J . Observation of individual DNA molecules undergoing gel electrophoresis. Science. 1989; 243(4888):203-6. DOI: 10.1126/science.2911733. View

10.

Bostock C . Parameters of field inversion gel electrophoresis for the analysis of pox virus genomes. Nucleic Acids Res. 1988; 16(10):4239-52. PMC: 336627. DOI: 10.1093/nar/16.10.4239. View

11.

Carle G, Frank M, Olson M . Electrophoretic separations of large DNA molecules by periodic inversion of the electric field. Science. 1986; 232(4746):65-8. DOI: 10.1126/science.3952500. View

12.

Turmel C, Brassard E, Slater G, Noolandi J . Molecular detrapping and band narrowing with high frequency modulation of pulsed field electrophoresis. Nucleic Acids Res. 1990; 18(3):569-75. PMC: 333463. DOI: 10.1093/nar/18.3.569. View

13.

Smisek D, Hoagland D . Electrophoresis of flexible macromolecules: evidence for a new mode of transport in gels. Science. 1990; 248(4960):1221-3. DOI: 10.1126/science.2349481. View

14.

Birren B, Simon M, Lai E . The basis of high resolution separation of small DNAs by asymmetric-voltage field inversion electrophoresis and its application to DNA sequencing gels. Nucleic Acids Res. 1990; 18(6):1481-7. PMC: 330515. DOI: 10.1093/nar/18.6.1481. View

15.

Holmes D, Stellwagen N . The electric field dependence of DNA mobilities in agarose gels: a reinvestigation. Electrophoresis. 1990; 11(1):5-15. DOI: 10.1002/elps.1150110103. View

16.

Stellwagen N . Anomalous electrophoresis of deoxyribonucleic acid restriction fragments on polyacrylamide gels. Biochemistry. 1983; 22(26):6186-93. DOI: 10.1021/bi00295a023. View

17.

Ansorge W, Barker R . System for DNA sequencing with resolution of up to 600 base pairs. J Biochem Biophys Methods. 1984; 9(1):33-47. DOI: 10.1016/0165-022x(84)90064-2. View

18.

Holmes D, Stellwagen N . 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. 1991; 12(9):612-9. DOI: 10.1002/elps.1150120903. View