Systematic study of field and concentration effects in capillary electrophoresis of DNA in polymer solutions.
A systematic study of the separation of double-stranded DNA in hydroxypropylcellulose (HPC) with a molecular mass of 10(6) was undertaken, using a variety of concentrations (from 0.1 to 1%) and different electric fields (from 6 to 540 V/cm). The data show that at high polymer concentrations ( > or = 0.4%) and low fields, the separation mechanism is similar to that occurring in gels. The results are in good agreement with theoretical models, and in particular with a recently proposed theory for gels with a pore size smaller than the persistence length of DNA. For more dilute solutions and high fields, however, the separation pattern cannot be explained by existing theories. The existence of an original mechanism was confirmed by the direct observation of the conformation of double-stranded DNA molecules in the polymer solution by fluorescence videomicroscopy. Practical conclusions for the capillary electrophoretic separation of duplex DNA are drawn.
Mitnik
L
L
Laboratoire de PhysicoChimie Théorique (URA CNRS 1382), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris, France.
Salomé
L
L
Viovy
J L
JL
Heller
C
C
D001583:Benzoxazoles; D002482:Cellulose / Q000031:analogs & derivatives; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D019075:Electrophoresis, Capillary; D007364:Intercalating Agents; D008433:Mathematics; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008956:Models, Chemical; D009690:Nucleic Acid Conformation; D011806:Quinolinium Compounds; D012996:Solutions
D001583:Benzoxazoles; D007364:Intercalating Agents; D011806:Quinolinium Compounds; D012996:Solutions; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; D002482:Cellulose; D004247:DNA; C008079:hydroxypropylcellulose