Аннотация
The possibility of separating appropriately labeled DNA fragments
using free-flow capillary electrophoresis was predicted a few years
ago based on simple theoretical arguments. Free-flow separation of
double-stranded DNA (dsDNA) fragments in the 100-1000 base range
was later demonstrated using a streptavidin label. In this article,
we now report that end-labeled free-flow electrophoresis (ELFSE)
can also be used to sequence single-stranded DNA (ssDNA). The first
100 bases of a DNA sequencing reaction were read without any sieving
matrix when fractionated streptavidin was added to the 5'-end of
the ssDNA fragments. These separations required only 18 min and did
not require coated capillaries. An analysis of the results indicates
that sample injection, analyte-wall interactions and thermal diffusion
are the limiting factors at this time. Extrapolating from our data,
we predict that several hundred bases could be sequenced in less
than 30 min with the proper conditions. ELFSE thus offers an attractive
potential alternative to polymer solutions for DNA sequencing in
capillaries and microchips.
Пользователи данного ресурса
Пожалуйста,
войдите в систему, чтобы принять участие в дискуссии (добавить собственные рецензию, или комментарий)