Separation and measurement of diffusion coefficients of linear and circular DNAs by flow field-flow fractionation

In this paper flow field-flow fractionation (flow FFF), an elution separation method, is utilized to separate and to measure the transnational diffusion coefficients D of a variety of linear and both single and double-stranded circular DNA chains in the molecular weight range M = (0.4 -4.8 x 10 supe...

Full description

Saved in:
Bibliographic Details
Published in:Macromolecules 1993-07, Vol.26 (14), p.3576-3588
Main Authors: Liu, Min Kuang, Giddings, J. Calvin
Format: Article
Language:English
Subjects:
550200 - Biochemistry
Applied sciences
BASIC BIOLOGICAL SCIENCES
Biological and medical sciences
CHEMICAL PROPERTIES
DNA
EQUATIONS
Exact sciences and technology
FRACTIONATION
Fundamental and applied biological sciences. Psychology
Miscellaneous
Molecular biophysics
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
Natural polymers
NUCLEIC ACIDS
ORGANIC COMPOUNDS
Physico-chemical properties of biomolecules
Physicochemistry of polymers
SEPARATION PROCESSES
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In this paper flow field-flow fractionation (flow FFF), an elution separation method, is utilized to separate and to measure the transnational diffusion coefficients D of a variety of linear and both single and double-stranded circular DNA chains in the molecular weight range M = (0.4 -4.8 x 10 super(6) Da. Equations for component retention times, band broadening, and resolution are given and compared with experimental results. The tradeoff between resolution and separation speed is discussed and experimentally realized. Overloading studies show that approximately equals 1 mu g of individual DNAs can be isolated per 10-20 min run; the procedure can be readily automated for repetitive runs. We find also that D is reasonably described by the simple form D = AM super(-b) over the 3-decade M range examined. Factors involved in the application of FFF to DNAs with M > 10 super(7) Da are discussed including shear degradation, transition to a steric mechanism of FFF, and use of condensed DNA. Severe overloading effects induced by chain entanglement rendered preliminary attempts unsuccessful, but future prospects for applying FFF to high-M DNA are found favorable.
ISSN:0024-9297
1520-5835
DOI:10.1021/ma00066a016