Mix of Sequencing Technologies for Sequence Closure: An Example

Abstract

s at 95 C followed by 30 s at 70 C. The sequencing gel was made of 8% Long Ranger gel solution (BMA, Rockland, ME, USA) and containing 7 M urea (Research Organics, Cleveland, OH, USA) and 1 TBE buffer (89 mM Tris, 89 mM boric acid, 2 mM EDTA . Na 2 [pH 8.4]). Three microliters of formamide loading dye (from the Amersham Pharmacia Biotech kit) were added to the samples after the cycle sequencing reaction. The loading membranes (sample loader) were from Genetic BioSystems (San Diego, CA, USA). The samples were heated for 2.5 min at 95 C and either immediately spotted onto the tabs of a preheated (95 C) membrane loader (hot sample application) or cooled down in ice-cold water bath before spotting (regular sample application).