DNA sequencing in HydroLink matrices: Extension of reading ability to > 600 nucleotides

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Abstract
All the systems for optimizing DNA sequencing published so far have introduced modifications regarding: (i) linearization of band migration via ionic strength gradients or wedge‐shaped gels; (ii) automatization of band reading via introduction of fluorescent probes; (iii) direct blotting analysis; (iv) pulsed electric fields and (v) discontinuous buffer systems. In all these systems, DNA sequence reading with an accuracy of ca. 98 % rarely exceeds a length of 350 bases. We have chosen, in order to increase the reading ability of a single gel, to manipulate the characteristics of the gel matrix. The Seq‐HydroLink gel formulation here reported allows optimal reading, from a single gel run, of at least 600 bases. In order to guarantee this reading ability in a single run, the upper and lower ends of the ladder are time‐resolved, i. e. the same sample is applied to the gel matrix at three different time intervals. The present system represents an increase of at least 30 % in reading ability as compared with any type of polyacrylamide gel formulation so far reported.