Electrotransfer of Long Ranger?Sequencing Gels Using a Methanol-TBE Buffer

摘要

Determining the primary structure of a nucleic acid is a multi-step process. The efficiency of each step must be maximized to gain the most sequence information. A critical and limiting step in this process is the electrophoresis of labeled or taggedfragments that terminate at a specific base. Historically, the electrophoresis has been performed using a polyacrylamide gel (PAG) matrix containing urea as a denaturant. The efficiency of the electrophoretic separation has been enhanced with the recentintroduction of the HydroLink~(R) Long Ranger~(TM) (HLR) gel matrix (AT Bio-chem, Malvern, PA, USA), which gives better separation of individual bands over an extended size range.