Micropreparative capillary electrophoresis of DNA by direct transfer onto a membrane.
We have developed a new technique for the collection of DNA fragments separated by capillary electrophoresis, by direct transfer from the capillary outlet to a positively charged membrane. Transfer and post-run detection of two different nonradioactively labeled DNA standards, ranging in size from 150 bp to 2 kbp and 120 bp to 23 kbp are presented, and discussed. Capillary electrophoresis with direct blotting presents several advantages over the blotting from gels: the separation is faster and requires less manual steps, the resolution is higher, and each DNA fragment is collected into a very concentrated spot on the membrane due to the small surface of the capillary outlet and to a design of the collection device inducing a refocusing of field lines across the hybridization membrane. Therefore, very small amounts of DNA (in the pg range) can be detected. This fraction collection makes further analysis of the sample possible, e.g. by hybridization, thus suppressing one of the major present limitations of the capillary electrophoresis technique for DNA analysis.
Magnúsdóttir
S
S
Institut Curie, Laboratoire de Physico-Chimie (UMR 168), Paris, France. soffia@curie.fr
Heller
C
C
Sergot
P
P
Viovy
J L
JL
D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D008567:Membranes, Artificial; D008832:Microchemistry
D008567:Membranes, Artificial; D004247:DNA