Glycerol-enhanced separation of DNA fragments in entangled solution capillary electrophoresis.
(Hydroxyalkyl)methylcellulose additives to borate buffers have been used to create physically entangled solutions for fractionation of biological molecules by high-performance capillary electrophoresis, i.e., ESCE. We have observed that addition of glycerol to an entangled solution containing (hydroxypropyl)methylcellulose (HPMC) in Tris-borate buffer markedly improved the separation of double-strand DNA molecules ranging from less than 100 bp to about 1 kb by ESCE. This range of DNA fragment sizes is well suited to many PCR-based studies. We attribute the improved ESCE performance to the borate present in the buffer system. Borate can form dimeric 1:2 borate:didiol complexes with both glycerol and HPMC, acting as a central linkage to allow an entangled solution with different pore size to form chemically.
Department of Molecular and Cell Biology, University of Aberdeen, U.K.
J
J
Cheng
K R
KR
Mitchelson
D000665:Ampholyte Mixtures / Q000737:chemistry; D001881:Borates / Q000737:chemistry; D005591:Chemical Fractionation / Q000379:methods; D004247:DNA / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods*; D005990:Glycerol / Q000737:chemistry*; D006801:Humans; D065347:Hypromellose Derivatives; D008747:Methylcellulose / Q000031:analogs & derivatives / Q000737:chemistry; D011108:Polymers / Q000737:chemistry
D000665:Ampholyte Mixtures; D001881:Borates; D011108:Polymers; C024617:polyol; D065347:Hypromellose Derivatives; D008747:Methylcellulose; D004247:DNA; D005990:Glycerol