High-Density Small-Volume Gel Loading Directly from Capillary Tubes
Author(s) -
Harold Evensen,
Deirdre R. Meldrum,
Channakhone Saenphimmachak,
Eric E. Dixon
Publication year - 1999
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/99275st05
Subject(s) - agarose , capillary action , loader , volume (thermodynamics) , materials science , sample (material) , chromatography , buffer (optical fiber) , capillary electrophoresis , composite material , chemistry , computer science , mechanical engineering , engineering , telecommunications , physics , quantum mechanics
A technique has been developed for high lane density loading of small-volume DNA samples in a horizontal agarose gel. This technique has been investigated with a simple hand-held tool that is made to couple to sample output from a new capillary-based sample automation system. The approach consists of piercing the gel with pressurized sample capillaries and relieving the pressure shortly before withdrawal. The pressurization prevents the capillary from aspirating the gel buffer and keeps the sample at the tip of the capillary, so that it may be sucked into the gel during withdrawal. This method is shown to be adequate for a wide range of DNA ladders and PCR-based screening. In addition to allowing smaller lanes and a higher lane density than is achievable with traditional well-forming techniques, it relaxes the need for well formation and the alignment of the sample loader with those wells, providing an easy, efficient means of loading agarose gels.
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