Influence of thickness of alkyl-silane coupling agent coating on separation of small DNA fragments in capillary gel electrophoresis

To simplify the process of coating capillaries with fused silica, we herein set out to develop a one-step procedure for coating capillaries to prevent electro-osmotic flow (EOF) during the separation of small DNA fragments. We selected a short capillary (total length = 15 cm; effective length = 7.5 cm) for use in a compact capillary gel electrophoresis (CGE) system. To develop a one-step coating procedure, we employed alkyltrimethoxysilane agents because they are cheap and can be easily acquired, in contrast to polyethylene glycol (PEG) silane coupling agents. We examined a 100-bp DNA Ladder sample using fused silica capillaries, which were coated with alkyltrimethoxysilane agents of five different molecular lengths (C4, C6, C8, C12, and C16). We found that a fused-silica capillary with C8 alkyltrimethoxysilane is optimal for separating small DNA samples.

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