ABSTRACT
Analysis of oligonucleotides with gel-filled capillary columns is a fast, efficient and automated way to check their purity. Gel-filled columns were optimized to separate oligonucleotides which were 20-50 nucleotides in length. The influence of sample size on resolution is discussed. A comparison between capillary gel electrophoresis and slab gel electrophoresis in terms of analysis time per sample is made. In addition to homopolymeric samples, the separation of heteropolymeric oligonucleotides with one nucleotide difference in size is presented and suggests the possibility of using capillary gel electrophoresis as a tool for DNA sequencing.
Subject(s)
Electrophoresis/methods , Oligonucleotides/analysisABSTRACT
Three immunogens with side chains of random amino acid sequence, poly (L Phe, l glu)-poly (DL Ala)--poly (L Lys) [(Phe, G)-A--L 223], poly (L Tyr, L Glu)-poly (DL Ala)--poly (L Lys) [(T, G)-A--L 509] and (T, G)-A-L 52, as well as two immunogens with side chains of defined amino acid sequences, GGT-A--L and TG-A--L, were sequenced using a Beckman automated sequenator. Despite the lack of a unique amino acid sequence for the amino terminus, reasonable results for the sequence studies were obtained using the Edman reaction. GGT-A--L and TG-A--L had 70% and 80% of their side chains respectively, with the desired sequence. The three compounds of random amino acid sequence were found to contain a large proportion of their A--L side chains unsubstituted. The side chains had a much greater probability of terminating in the aromatic amino acid than in the glutamic acid. The distribution of the length of side chains and their amino acid sequences was completely heterogeneous.
