Photo-polymerized cross-linked polyacrylamide gels are becoming increasingly important for use in micro-fabricated DNA electrophoresis systems because they allow a concentrated sieving matrix to be precisely positioned at any location within a complex micro-channel network. The rate of photo-initiation in the free radical gel polymerization reaction, however, can exert a strong influence on the resulting gel structure. Experimental data on separation resolution of single stranded DNA (ssDNA) in photoinitiated polyacrylamide gels is very sparse. In this study, we investigate the performance of ssDNA electrophoresis in an ALF Express automated DNA sequencer using various photo-initiation chemistries. Cross-linked polyacrylamide gels with concentrations ranging from 6 to 12 %T were prepared using riboflavin, methylene blue, irgacure 651 and ReproGel (AP Biotech) photo-initiators. Separation resolution is compared with that attained in cross-linked polyacrylamide gels prepared using conventional chemical initiators (e.g. ammonium persulfate/ Tetramethylethylenediamine) in order to determine the polymerization conditions necessary for optimum performance.
Photo-polymerized cross-linked polyacrylamide gels are becoming increasingly important for use in micro-fabricated DNA electrophoresis systems because they allow a concentrated sieving matrix to be precisely positioned at any location within a complex micro-channel network. The rate of photo-initiation in the free radical gel polymerization reaction, however, can exert a strong influence on the resulting gel structure. Experimental data on separation resolution of single stranded DNA (ssDNA) in photoinitiated polyacrylamide gels is very sparse. In this study, we investigate the performance of ssDNA electrophoresis in an ALF Express automated DNA sequencer using various photo-initiation chemistries. Cross-linked polyacrylamide gels with concentrations ranging from 6 to 12 %T were prepared using riboflavin, methylene blue, irgacure 651 and ReproGel (AP Biotech) photo-initiators. Separation resolution is compared with that attained in cross-linked polyacrylamide gels prepared using conventional chemical initiators (e.g. ammonium persulfate/ Tetramethylethylenediamine) in order to determine the polymerization conditions necessary for optimum performance.
