A DNA sequencing system based on the use of a novel set of four chain-terminating dideoxynucleotides, each carrying a different chemically tuned succinylfluorescein dye distinguished by its fluorescent emission is described. Avian myeloblastosis virus reverse transcriptase is used in a modified dideoxy DNA sequencing protocol to produce a complete set of fluorescence-tagged fragments in one reaction mixture. These DNA fragments are resolved by polyacrylamide gel electrophoresis in one sequencing lane and are identified by a fluorescence detection system specifically matched to the emission characteristics of this dye set. A scanning system allows multiple samples to be run simultaneously and computer-based automatic base sequence identifications to be made. The sequence analysis of M13 phage DNA made with this system is described.
FOUR PLATES (THIRTY FIGURES) AUTHOR'S ABSTRACTDuring the quiescent fourth larval instar the larva of Drosophillc becomes loosened from the puparium and takes on the form of a pupa . A large gas bubble is formed within the b2dAt the end of this stadmm. which lasts 111 hours a t 25 d it upates shedding the prepupal cuticle . The gas bubble is used as a meqhanistn to separ;te txe cutiile from its hypodermis . Practically all of the tissues undergo hlstolysls and are replaced by imaginal structures derived from specialized cells . Some of the muscles persist longer than the general mass and perform special functions in the pupa . The sequence of histolytic and histogenetic changes is accurately timed at a constant temperature . during this prepupal period .
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